A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine, rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines

Abstract

The present invention provides a compound represented by the formula:

wherein R1 is a C1-4alkyl; R2 is (1) a 5- to 7-membered nitrogen-containing heterocyclic group which may have a substituent selected from the group consisting of (1′) a halogen, (2′) a hydroxy group, (3′) a C1-4alkyl and (4′) a C1-4alkoxy, (2) a phenyl which may have a substituent selected from the group consisting of (1′) a halogen, (2′) a C1-4alkoxy-C1-4alkyl, (3′) a mono-C1-4alkyl-carbamoyl-C1-4alkyl, (4′) a C1-4alkoxy and (5′) a mono-C1-4alkylcarbamoyl-C1-4alkoxy, or the like; R3 is a C1-4alkyl; R4 is a C1-4alkoxy, or the like; n is an integer of 1 to 4; or a salt thereof, as a thienopyrimidine compound having gonadotropin-releasing hormone antagonistic activity.

Description

This application is the National Phase filing of International Patent Application No. PCT/JP2004/000741, filed January 28, 2004.

Secretion of anterior pituitary hormones undergoes feedback control by peripheral hormones secreted from target organs of the respective hormones and by secretion-regulating hormones from the hypothalamus, which is the upper central organ of the anterior lobe of the pituitary (hereinafter, these hormones are collectively called “hypothalamic hormones” in this specification). Presently, as hypothalamic hormones, the existence of nine kinds of hormones including, for example, thyrotropin releasing hormone (TRH), and gonadotropin releasing hormone [GnRH, sometimes called as LH-RH (luteinizing hormone releasing hormone)] has been confirmed. These hypothalamic hormones are believed to show their actions via the receptors which are considered to exist in the anterior lobe of the pituitary, and efforts to find the receptor-gene expression specific to these hormones, including cases of human, have been made. Accordingly, antagonists or agonists specifically and selectively acting on these receptors should control the action of the hypothalamic hormone and the secretion of anterior pituitary hormone. As a result, such antagonists or agonists are expected to prevent or treat anterior pituitary hormone dependent diseases.

Peptide compounds pose a large number of problems to be resolved with respect to oral absorbability, dosage form, dose volume, drug stability, sustained action, metabolic stability etc. There is strong demand for an oral GnRH antagonist, especially one based on a non-peptide compound, that has excellent therapeutic effect on hormone-dependent cancers, e.g., prostatic cancer, endometriosis, precocious puberty etc., that does not show transient hypophysial-gonadotropic action (acute action) and that has excellent oral absorbability.

DISCLOSURE OF INVENTION

We, the present inventors, have conducted various investigations, and as a result, have synthesized the following novel compound represented by the formula [hereinafter sometimes referred to briefly as Compound (I)]:

wherein

R1 is a C1-4alkyl; R2 is (1) a C1-6alkyl which may have a substituent selected from the group consisting of (1′) a hydroxy group, (2′) a C1-4 alkoxy, (3′) a C1-4alkoxy-carbonyl, (4′) a di-C1-4alkyl-carbamoyl, (5′) a 5- to 7-membered nitrogen-containing heterocyclic group, (6′) a C1-4alkyl-carbonyl and (7′) a halogen,

(2) a C3-8cycloalkyl which may have (1′) a hydroxy group or (2′) a mono-C1-4alkyl-carbonylamino, (3) a 5- to 7-membered nitrogen-containing heterocyclic group which may have a substituent selected from the group consisting of (1′) a halogen, (2′) a hydroxy group, (3′) a C1-4alkyl and (4′) a C1-4alkoxy, (4) a phenyl which may have a substituent selected from the group consisting of (1′) a halogen, (2′) a C1-4alkoxy-C1-4alkyl, (3′) a mono-C1-4alkyl-carbamoyl-C1-4alkyl, (4′) a C1-4alkoxy and (5′) a mono-C1-4 alkylcarbamoyl-C1-4alkoxy or (5) a C1-4alkoxy; R3 is a C1-4alkyl; R4 is (1) a hydrogen atom, (2) a C1-4alkoxy, (3) a C6-10 aryl, (4) a N—C1-4alkyl-N—C1-4alkylsulfonylamino, (5) a hydroxy group or (6) a 5- to 7-membered nitrogen-containing heterocyclic group which may have a substituent selected from the group consisting of (1′) oxo, (2′) a C1-4alkyl, (3′) a hydroxy-C1-4alkyl, (4′) a C1-4alkoxy-carbonyl, (5′) a mono-C1-4alkyl-carbamoyl and (6′) a C1-4alkylsulfonyl; n is an integer of 1 to 4; provided that when R2 is a phenyl which may have a substituent, R4 is a 5- to 7-membered nitrogen-containing heterocyclic group which may have a substituent selected from the group consisting of (1) oxo, (2) a hydroxy-C1-4alkyl, (3) a C1-4alkoxy-carbonyl, (4) a mono-C1-4alkyl-carbamoyl and (5) a C1-4alkylsulfonyl; or a salt thereof; which is characterized by having 3-C1-4alkoxyureido on the para-position of its phenyl group at the six position of the thieno[2,3-d]pyrimidine skeleton. And we also have found that Compound (I) has an unexpected, excellent GnRH-antagonizing activity, especially strong antagonistic activity, based upon the above specific chemical structure, and extremely low toxicity and is therefore satisfactory as a medicine having GnRH-antagonizing activity, and developed the present invention based on these findings.

Accordingly, the present invention relates to:

[1] A compound of the formula:

wherein

R1 is a C1-4alkyl;

R2 is

(1) a C1-6alkyl which may have a substituent selected from the group consisting of (1′) a hydroxy group, (2′) a C1-4alkoxy, (3′) a C1-4alkoxy-carbonyl, (4′) a di-C1-4alkyl-carbamoyl, (5′) a 5- to 7-membered nitrogen-containing heterocyclic group, (6′) a C1-4alkyl-carbonyl and (7′) a halogen,

(2) a C3-8cycloalkyl which may have (1′) a hydroxy group or (2′) a mono-C1-4alkyl-carbonylamino,

(3) a 5- to 7-membered nitrogen-containing heterocyclic group which may have a substituent selected from the group consisting of (1′) a halogen, (2′) a hydroxy group, (3′) a C1-4alkyl and (4′) a C1-4alkoxy,

(4) a phenyl which may have a substituent selected from the group consisting of (1′) a halogen, (2′) a C1-4alkoxy-C1-4alkyl, (3′) a mono-C1-4alkyl-carbamoyl-C1-4alkyl, (4′) a C1-4alkoxy and (5′) a mono-C1-4alkylcarbamoyl-C1-4alkoxy, or

(5) a C1-4alkoxy;

R3 is a C1-4alkyl;

R4 is

(1) a hydrogen atom,

(2) a C1-4alkoxy,

(3) a C6-10aryl,

(4) a N—C1-4alkyl-N—C1-4alkylsulfonylamino,

(5) a hydroxyl group, or

(6) a 5- to 7-membered nitrogen-containing heterocyclic group which may have a substituent selected from the group consisting of (1′) oxo, (2′) a C1-4alkyl, (3′) a hydroxy-C1-4alkyl, (4′) a C1-4alkoxy-carbonyl, (5′) a mono-C1-4alkyl-carbamoyl and (6′) a C1-4alkylsulfonyl;

n is an integer of 1 to 4;
provided that when R2 is a phenyl which may have a substituent, R4 is a 5- to 7-membered nitrogen-containing heterocyclic group which may have a substituent selected from the group consisting of (1) oxo, (2) a hydroxy-C1-4alkyl, (3) a C1-4alkoxy-carbonyl, (4) a mono-C1-4alkyl-carbamoyl and

(5) a C1-4alkylsulfonyl; or a salt thereof;

[2] A compound as defined in [1] above, wherein

R2 is

(1) a C1-4alkyl which may have a substituent selected from the group consisting of (1′) a hydroxy group, (2′) a C1-4alkoxy, (3′) a C1-4alkoxy-carbonyl, (4′) a di-C1-4alkyl-carbamoyl and (5′) a 5 to 7-membered nitrogen-containing heterocyclic group,

(2) a C3-8cycloalkyl which may have (1′) a hydroxy group or (2′) a mono-C1-4alkyl-carbonylamino,

(3) a 5- to 7-membered nitrogen-containing heterocyclic group which may have a substituent selected from the group consisting of (1′) a halogen,

(2′) a hydroxy group, (3′) a C1-4alkyl and (4′) a C1-4alkoxy,

(4) a phenyl which may have a substituent selected from the group consisting of (1′) a halogen, (2′) a C1-4alkoxy-C1-4alkyl, (3′) a mono-C1-4alkyl-carbamoyl-C1-4alkyl and (4′) a mono-C1-4alkylcarbamoyl-C1-4alkoxy, or

(5) a C1-4alkoxy;

R4 is

(1) a C1-4alkoxy,

(2) a C6-10aryl,

(3) a N—C1-4alkyl-N—C1-4alkylsulfonylamino or

(4) a 5- to 7-membered nitrogen-containing heterocyclic group which may have a substituent selected from the group consisting of (1′) oxo, (2′) a hydroxy-C1-4alkyl, (3′) a C1-4alkoxy-carbonyl, (4′) a mono-C1-4alkyl-carbamoyl and (5′) a C1-4alkylsulfonyl;

[3] A compound as defined in [1] above, wherein R1 is methyl;

[4] A compound as defined in [1] above, wherein R2 is a 5- to 7-membered nitrogen-containing heterocyclic group which may have a substituent selected from the group consisting of (1) a halogen, (2) a hydroxy group, (3) a C1-4alkyl and (4) a C1-4alkoxy;

[5] A compound as defined in [1] above, wherein R3 is methyl;

[6] A compound as defined in [1] above, wherein R4 is a C1-4alkoxy;

[7] A compound as defined in [1] above, wherein n is 2;

[8] A compound as defined in [1] above, wherein R3 is methyl, R4 is a hydrogen atom and n is 1;

[9] N-(4-(1-(2,6-difluorobenzyl)-5-(((2-methoxyethyl)(methyl)amino)methyl)-2,4-dioxo-3-(2-pyridinyl)-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea, N-(4-(1-(2,6-difluorobenzyl)-5-(((2-ethoxyethyl)(methyl)amino)methyl)-2,4-dioxo-3-(2-pyridinyl)-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea, N-(4-(1-(2,6-difluorobenzyl)-5-((dimethylamino)methyl)-3-(6-methoxy-3-pyridazinyl)-2,4-dioxo-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea or N-(4-(1-(2,6-difluorobenzyl)-5-((dimethylamino)methyl)-3-(6-methoxypyridin-3-yl)-2,4-dioxo-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea, or a salt thereof;

[10] N-(4-(5-(((2-methoxyethyl)methylamino)methyl)-1-(2,6-difluorobenzyl)-1,2,3,4-tetrahydro-2,4-dioxo-3-(4-methoxyphenyl)thieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea or N-(4-(1-(2,6-difluorobenzyl)-5-((dimethylamino)methyl)-3-(4-methoxyphenyl)-2,4-dioxo-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea, or a salt thereof;

[11] A prodrug of the compound as defined in [1] above;

[12] A pharmaceutical which comprises the compound as defined in [1] above or a prodrug thereof;

[13] A pharmaceutical as defined in [12] above, which is a gonadotropin-releasing hormone antagonist;

[14] A pharmaceutical as defined in [12] above, which is for preventing or treating a sex hormone dependent disease;

[16] A method for antagonizing gonadotropin-releasing hormone, which comprises administering an effective amount of the compound as defined in [1] above to a mammal;

[17] Use of the compound as defined in [1] above for manufacturing a pharmaceutical composition for antagonizing gonadotropin-releasing hormone;

[18] A compound of the formula:

wherein

Ra is (1) a hydrogen atom, (2) an aryl group which may have 1 to 5 substituents selected from the group consisting of (i) a halogen, (ii) nitro, (iii) cyano, (iv) amino, (v) a carboxyl group which may be esterified or amidated, (vi) an alkylenedioxy, (vii) an alkyl, (viii) an alkoxy, (ix) an alkylthio, (x) an alkylsulfinyl and (xi) an alkylsulfonyl, (3) a cycloalkyl group which may have a substituent or (4) a heterocyclic group which may have a substituent; Rb is a nitrogen-containing heterocyclic group which may have a substituent; Rc is an amino group which may have a substituent; Rd is an aryl group which may have a substituent; p is an integer of 0 to 3; q is an integer of 0 to 3; or a salt thereof; and so forth.

The definition of each term is described in the following paragraphs.

Examples of the “C1-4alkyl” include a linear C1-4alkyl (e.g. methyl, ethyl, propyl, butyl, and the like), a branched C3-4alkyl (e.g., isopropyl, isobutyl, sec-butyl, tert-butyl, and the like), and the like.

Examples of the “C1-6alkyl” include a linear C1-6alkyl (e.g. methyl, ethyl, propyl, butyl, pentyl, hexyl and the like), a branched C3-6alkyl (e.g., isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl and the like), and the like.

Examples of the “C1-4alkoxy” include a linear C1-4alkoxy (e.g. methoxy, ethoxy, propoxy, butoxy, and the like), a branched C3-4alkoxy (e.g., isopropoxy, isobutoxy, sec-butoxy, tert-butoxy, and the like), and the like.

Examples of the “C1-4alkoxy-carbonyl” include methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, sec-butoxycarbonyl, tert-butoxycarbonyl, and the like.

Examples of the “di-C1-4alkyl-carbamoyl” include dimethylcarbamoyl, diethylcarbamoyl, dipropylcarbamoyl, diisopropylcarbamoyl, N-ethyl-N-methylcarbamoyl, and the like.

Examples of the “mono-C1-4alkyl-carbamoyl” include methylcarbamoyl, ethylcarbamoyl, propylcarbamoyl, isopropylcarbamoyl, butylcarbamoyl, isobutylcarbamoyl, sec-butylcarbamoyl, tert-butylcarbamoyl, and the like.

Examples of the “C1-14alkylsulfonyl” include methylsulfonyl, ethylsulfonyl, propylsulfonyl, isopropylsulfonyl, butylsulfonyl, isobutylsulfonyl, sec-butylsulfonyl, tert-butylsulfonyl, and the like.

As R1, methyl and ethyl are preferable, and especially methyl is preferable.

As R2, 5- to 7-membered nitrogen-containing heterocyclic group which may have a substituent selected from the group consisting of (1) a halogen, (2) a hydroxy group, (3) a C1-4alkyl and (4) a C1-4alkoxy is preferable. Among them, pyridyl (pyridin-2-yl, pyridin-3-yl, pyridin-4-yl), which may have a substituent selected from the group consisting of (1) a halogen, (2) a hydroxy group, (3) a C1-4alkyl and (4) a C1-4alkoxy is more preferable. Especially, unsubstituted pyridin-2-yl is preferable.

As R3, methyl and ethyl are preferable. Especially, methyl is preferable.

As R4, a C1-4alkoxy is preferable. Especially, methoxy and ethoxy are preferable.

As n, 1 or 2 is preferable. Especially, 2 is preferable.

Preferable examples of the combination of R3, R4 and n, includes the case that R3 is methyl, R4 is a hydrogen atom and n is 1.

For example, Compound (I) can be produced according to the following production methods. Compounds illustrated in the following reaction schemes include their salts. Examples of the salts include the same salts as the salts of Compound (I), etc. Compounds (I)-(IV) illustrated in the following reaction schemes may be formed into the acceptable salts depending on the reaction conditions.

In the above formula, L is a leaving group, and other symbols are the same as defined above.

Examples of “leaving group” represented by L are a halogen atom, C1-4alkylsulfonyloxy which may have a halogen atom, and the like.

Examples of “C1-4alkylsulfonyloxy which may have a halogen atom” are methanesulfonyloxy, ethanesulfonyloxy, trifluoromethanesulfonyloxy, and the like.

Compound (II) can be produced in any per se known manner, for example, according to the methods disclosed in JP-A-2001-278884, WO 00/56739 or analogous methods thereto.

For example, Compound (I) can be produced by reacting Compound (II) and a compound represented by the formula: R4—(CH2)n-L. This reaction is preferably carried out in the presence of a base.

The amount of the compound represented by the formula: R4—(CH2)n-L in the reaction of Compound (II) and the compound represented by the formula: R4—(CH2)n—L is about 1 to about 3 moles per 1 mole of Compound (II). The amount of a base is about 1 to about 3 moles per 1 mole of Compound (II).

This reaction is usually carried out in a solvent inert to the reaction. Examples of “solvent” are an ether (e.g., diethyl ether, dioxane, dimethoxyethane, tetrahydrofuran, and the like), an aromatic hydrocarbon (e.g., benzene, toluene, and the like), an amide (e.g., dimethylformamide, dimethylacetamide, and the like), a halogenated hydrocarbon (e.g., chloroform, dichloromethane, and the like), and the like.

The reaction temperature is usually about 0 to about 150° C., preferably about 50 to about 80° C. The reaction time is usually about 1 to about 24 hours.

In the above formula, R′ is a hydrogen atom or a C1-4alkyl; R″ is a C1-4alkyl; and the other symbols are as defined above.

Examples of the C1-4alkyl represented by R′ and R″ are a linear C1-4alkyl (e.g., methyl, ethyl, propyl, butyl, and the like), a branched C3-4alkyl (e.g., isopropyl, isobutyl, sec-butyl, tert-butyl, and the like), and the like.

Compound (II) can be produced in any per se known manner, for example, by reacting p-nitrophenylacetone with a cyanoacetic ester compound and sulphur [e.g., Chem. Ber., 99, 94-100(1966)] followed by subjecting the obtained 2-amino-4-methyl-5-(4-nitrophenyl)thiophene to the methods disclosed in JP-A-9-169768, WO 96/24597 or analogous methods thereto.

1) When R′ is hydrogen atom, Compound (I) can be produced by reacting Compound (III) with a compound represented by the formula: R2—NH2 or a salt thereof in the presence of a condensing agent, to obtain Compound (IV), following by subjecting to cycization.

The amount of “condensing agent” is about 1 to about 3 moles per 1 mole of Compound (III).

This reaction is advantageously carried out in a solvent inert to the reaction.

Examples of the solvent are an alcohol (e.g., ethanol, methanol, and the like), an aromatic hydrocarbon (e.g., benzene, toluene, and the like), an amide (e.g., dimethylformamide, dimethylacetamide, and the like), a halogenated hydrocarbon (e.g., chloroform, dichloromethane, and the like), and so the like.

The reaction temperature is usually about 0 to about 150° C., preferably about 0 to 25° C. The reaction time is usually about 1 to about 36 hours.

The product as produced in the manner mentioned above may be applied to the next reaction while it is still crude in the reaction mixture, or may be isolated from the reaction mixture in any ordinary manner.

The amount of “base” is about 2 to about 20 moles, preferably about 5 to about 12 mole per 1 mole of Compound (IV).

This reaction is usually carried out in a solvent inert to the reaction.

Examples of the solvent are an alcohol (e.g., ethanol, methanol, and the like), an aromatic hydrocarbon (e.g., benzene, toluene, and the like), an amide (e.g., dimethylformamide, dimethylacetamide, and the like), a halogenated hydrocarbon (e.g., chloroform, dichloromethane, and the like), and the like.

The reaction temperature is usually about 0 to 150° C., preferably room temperature (about 15 to 25° C.). The reaction time is usually about 1 to 36 hours.

2) When R′ is an alkyl group, Compound (I) can be produced by reacting Compound (III) with an activated R2—NH2.

The activated R2—NH2 can be produced in any per se known manner, for example, by reacting an organo-aluminum reagent with R2—NH2 in a solvent inert to the reaction.

Examples of “organo-aluminum reagent” are trimethyl aluminum, dimethyl aluminum chloride, and the like; and a solution including them, and the like.

The amount of “organo-aluminum reagent” is about 1 to about 5 moles, preferably about 1 mole per 1 mole of R2—NH2.

Examples of the solvent are a halogenated hydrocarbon (e.g., chloroform, dichloromethane, and the like).

The reaction temperature is usually about 0 to about 150° C., preferably about 0 to 25° C. The reaction time is usually about 1 to about 6 hours.

The cyclization can be carried out by reacting Compound (III) with an activated R2—NH2 to obtain Compound (I).

The amount of “Compound (III)” is about 1/5 volume of a mixture of R2—NH2 and the organo-aluminum reagent.

This reaction is usually carried out in a solvent inert to the reaction.

Such solvent is the same as those used in the reaction to obtain an activated R2—NH2.

The reaction temperature is usually about 0 to about 150° C., preferably about 0 to 25° C. The reaction time is usually about 1 to about 48 hours.

Compound (I) may be isolated and purified by per se known means of separation such as recrystallization, distillation and chromatography, and the like.

When Compound (I) is obtained in free form, it can be converted to a salt by per se known methods or methods analogous thereto. When Compound (I) is obtained in salt form, it can be converted to the free form or another salt by per se known methods or methods analogous thereto.

Compound (I) may be a hydrate or a non-hydrate. The hydrate is exemplified by monohydrate, sesquihydrate and dihydrate.

When Compound (I) is obtained as a mixture of optically active configurations, it can be resolved into the (R)- and (S)-forms by the conventional optical resolution techniques.

Compound (I) can be used as a prodrug. The prodrug of Compound (I) or a salt thereof means a compound which is converted to Compound (I) of the present invention under physiological conditions or with a reaction due to an enzyme, a gastric acid, and the like in the living body, that is, a compound which is converted to Compound (I) of the present invention with oxidation, reduction, hydrolysis, and the like according to an enzyme; a compound which is converted to Compound (I) of the present invention with gastric acid, etc. The prodrug for Compound (I) may for example be a compound obtained by subjecting an amino group in Compound (I) to an acylation, alkylation or phosphorylation (e.g., a compound obtained by subjecting an amino group in Compound (I) or to an eicosanoylation, alanylation, pentylaminocarbonylation, (5-methyl-2-oxo-1,3-dioxolen-4-yl)methoxycarbonylation, tetrahydrofuranylation, pyrrolidylmethylation, pivaloyloxymethylation and tert-butylation, etc.); a compound obtained by subjecting a hydroxy group in Compound (I) to an acylation, alkylation, phosphorylation or boration (e.g., a compound obtained by subjecting an hydroxy in Compound (I) to an acetylation, palmitoylation, propanoylation, pivaloylation, succinylation, fumarylation, alanylation, dimethylaminomethylcarbonylation, etc.). Any of these compounds can be produced from Compound (I) by a method known per se.

A prodrug of Compound (I) may also be one which is converted into Compound (I) under a physiological condition, such as those described in “IYAKUHIN no KAIHATSU (Development of Pharmaceuticals)”, Vol. 7, Design of Molecules, p. 163-198, Published by HIROKAWA SHOTEN (1990).

Compound (I) may be labeled with an isotope (e.g., 3H, 14C, 35S) and the like.

In the reaction described above, a starting compound having an amino group, a carboxy group or a hydroxy group as its substituent may be present as a compound in which a protective group employed ordinarily in a peptide chemistry has been introduced into such a substituent, and an intended compound can be obtained by deprotection if necessary after the reaction.

A protective group for an amino group may for example be an optionally substituted C1-6alkyl-carbonyl (e.g., acetyl, propionyl, and the like), formyl, phenylcarbonyl, a C1-6alkyloxycarbonyl (e.g., methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, and the like), phenyloxycarbonyl, a C7-14 aralkyloxy-carbonyl (e.g., benzyloxycarbonyl, 9-fluorenylmethoxycarbonyl, and the like), trityl, phthaloyl and the like. Its substituent may for example be a halogen atom (e.g., fluorine, chlorine, bromine and iodine), a C1-6alkylcarbonyl (e.g., acetyl, propionyl, butyryl, and the like), nitro and the like, and the number of the substituents may be 1 to 3.

A protective group for a carboxy may for example be an optionally substituted C1-6alkyl (e.g., methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, and the like), phenyl, trityl, silyl and the like. Its substituent may for example be a halogen atom (e.g., fluorine, chlorine, bromine and iodine), a C1-6alkylcarbonyl (e.g., acetyl, propionyl, butyryl, and the like), formyl, nitro, and the number of the substituents may be 1 to 3.

A protective group for a hydroxy group may for example be an optionally substituted C1-6alkyl (e.g., methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, and the like), phenyl, a C7-10 aralkyl (e.g., benzyl, and the like), a C1-6alkylcarbonyl (e.g., acetyl, propionyl, and the like), formyl, phenyloxycarbonyl, a C7-10 aralkyloxycarbonyl (e.g., benzyloxycarbonyl, and the like), tetrahydropyranyl, tetrahydrofuranyl, silyl and the like. Its substituent may for example be a halogen atom (e.g., fluorine, chlorine, bromine and iodine), a C1-6alkyl, phenyl, a C7-11 aralkyl, nitro, and the like, and the number of the substituents may be 1 to 4.

The method for introducing and removing the protective group is demonstrated in accordance with a known method or analogous method thereof (e.g., the method described in Protective Groups in Organic Chemistry (J. F. W. McOmie et al, Plenum Press)). A deprotection method may be a treatment with an acid, base, reduction, UV, hydrazine, phenylhydrazine, sodium N-methyldithiocarbamate, tetrabutylammonium fluoride, palladium acetate and the like.

The compound of the present invention can therefore be safely used in a mammal (e.g., human, monkey, bovine, horse, dog, cat, rabbit, rat, mouse, etc.) for the preventing and/or treating diseases depending on male or female hormones, diseases due to excess of these hormones, etc., by suppressing gonadotropin secretion with its GnRH receptor-antagonizing action to control plasma sex hormone concentrations.

For example, the compound of the present invention is useful for preventing and/or treating sex hormone-dependent cancers (e.g., prostatic cancer, uterine cancer, breast cancer, pituitary tumor, etc.), bone metastasis of sex hormone-dependent cancer, prostatic hypertrophy, hysteromyoma, endometriosis, metrofibroma, precocious puberty, amenorrhea, premenstrual syndrome, dysmenorrhea, multilocular ovary syndrome, polycystic ovary syndrome, acne, alopecia, Alzheimer's disease (Alzheimer's disease, senile dementia of Alzheimer type and a mixed type thereof), and the like. The compound of the present invention is also useful for the regulation of reproduction in males and females (e.g., pregnancy regulators, menstruation cycle regulators, etc.). The compound of the present invention can be also used as a male or female contraceptive, or as a female ovulation inducer. Based on its rebound effect after withdrawal, the compound of the present invention can be used to treat infertility. And the compound of this invention can be used as an agent for preventing and/or treating benign or malignant tumor which is hormone independent and LH-RH sensitive. And the compound of the present invention can be used as an agent for preventing and/or treating irritable bowel syndrome and for preventing postoperative recurrence of sex hormone-dependent cancer (an agent for preventing postoperative recurrence of prostatic cancer; an agent for preventing postoperative recurrence of breast cancer or ovarian cancer in the condition before or after menopause; especially, an agent for preventing postoperative recurrence of breast cancer or ovarian cancer in the condition before menopause).

In addition, the compound of the present invention is useful for regulation of animal estrus, improvement of meat quality and promotion of animal growth in the field of animal husbandry. The compound of the present invention is also useful as a fish spawning promoter.

The compound of the present invention can be also used to suppress the transient rise in plasma testosterone concentration (flare phenomenon) observed in administration of a GnRH super-agonist such as leuprorelin acetate. The compound of the present invention can be used in combination with a GnRH super-agonist such as leuprorelin acetate, gonadorelin, buserelin, triptorelin, goserelin, nafarelin, histrelin, deslorelin, meterelin, lecirelin, and the like. Among others, preferred is leuprorelin acetate.

It is also beneficial to use the compound of the present invention in conjunction with at least one member selected from the steroidal or nonsteroidal antiandrogen agent or antiestrogen agent, chemotherapeutic agent, GnRH antagonistic peptide, α-reductase inhibitor, α-receptor inhibitor, aromatase inhibitor, 17β-hydroxysteroid dehydrogenase inhibitor, adrenal androgen production inhibitor, kinase inhibitor, drug for hormone therapy, and drug inhibiting cell growth factor or its receptor, among others.

As a further class of drugs inhibiting the cell growth factor or its receptor includes HER2 inhibitors. The HER2 inhibitor may be any substance that inhibits the activity of HER2 (e.g., phosphorylating activity), thus including an antibody, a low-molecular weight compound (synthetic or natural product), an antisense, an HER2 ligand, heregulin, and any of them as partially modified or mutated in structure. Moreover, it may be a substance which inhibits HER2 activity by inhibiting HER2 receptor (e.g. HER2 receptor antibody). The low molecular weight compound having HER2 inhibiting activity includes, for example, the compounds described in WO 98/03505, namely 1-[3-[4-[2-((E)-2-phenylethenyl)-4-oxazolylmethoxy]phenyl]propyl]-1,2,4-triazole and the like.

The administration mode of the compound of the present invention and a concomitant medicament are not particularly limited, provided that the compound of the present invention and the concomitant medicament are combined upon administration. Such an administration mode may for example be (1) an administration of a single formulation obtained by formulating the compound of the present invention and a concomitant medicament simultaneously, (2) a simultaneous administration via an identical route of two formulations obtained by formulating the compound of the present invention and a concomitant medicament separately, (3) a sequential and intermittent administration via an identical route of two formulations obtained by formulating the compound of the present invention and a concomitant medicament separately, (4) a simultaneous administration via different routes of two formulations obtained by formulating the compound of the present invention and a concomitant medicament separately, (5) a sequential and intermittent administration via different routes of two formulations obtained by formulating the compound of the present invention and a concomitant medicament separately (for example, the compound of the present invention followed by concomitant medicament, or inverse order) and the like.

When the compound of the present invention is used as a preventing and/or treating agent for the above-mentioned diseases or used in the field of animal husbandry or fishery, it can be administered orally or non-orally, as formulated with a pharmaceutically acceptable carrier, normally in the form of solid preparations such as tablets, capsules, granules and powders for oral administration, or in the form of intravenous, subcutaneous, intramuscular or other injections, suppositories or sublingual tablets for non-oral administration. It may also be sublingually, subcutaneously, intramuscularly or otherwise administered in the form of sustained-release preparations of sublingual tablets, microcapsules, etc. Depending on symptom severity; subject age, sex, weight and sensitivity; duration and intervals of administration; property, dispensing and kind of pharmaceutical preparation; kind of active ingredient etc., daily dose is not subject to limitation. For use in the treatment of the above-described sex hormone-dependent cancers (e.g., prostatic cancer, uterine cancer, breast cancer, pituitary tumor, etc.), prostatic hypertrophy, hysteromyoma, endometriosis, precocious puberty etc., daily dose is normally about 0.01 to 30 mg, preferably about 0.02 to 10 mg, and more preferably 0.1 to 10 mg, especially preferably 0.1 to 5 mg per kg weight of mammal, normally in 1 to 4 divided dosages.

The above doses of the active ingredient (the compound of the present invention) for oral administration are applicable to the use of the compound of the present invention in the field of animal husbandry or fishery. Daily dose is about 0.01 to 30 mg, preferably about 0.1 to 10 mg, per kg weight of subject organism, normally in 1 to 3 divided dosages.

In the pharmaceutical composition of the present invention, the amount of Compound (I) is 0.01 to 100% by weight or so of the total weight of the composition.

The above pharmaceutically acceptable carriers are various organic or inorganic carrier substances in common use as pharmaceutical materials, including excipients, lubricants, binders and disintegrants for solid preparations; solvents, dissolution aids, suspending agents, isotonizing agents, buffers and soothing agents for liquid preparations; and the like. Other pharmaceutical additives such as preservatives, antioxidants, coloring agents and sweetening agents may be used as necessary.

By adding suspending agents, dissolution aids, stabilizers, isotonizing agents, preservatives, and the like, the compound of the present invention can be prepared as an intravenous, subcutaneous or intramuscular injection by a commonly known method. In such cases, the compound of the present invention can be freeze-dried as necessary by a commonly known method. In administration to humans, for example, the compound of the present invention can be safely administered orally or non-orally as such or as a pharmaceutical composition prepared by mixing it with a pharmacologically acceptable carrier, excipient and diluent selected as appropriate.

These preparations can be produced by commonly known methods in common use for pharmaceutical making processes.

An injection can be produced by, for example, preparing the compound of the present invention as an aqueous injection along with a dispersing agent (e.g., Tween 80 (produced by Atlas Powder Company, USA), HCO 60 (produced by Nikko Chemicals Co., Ltd.), polyethylene glycol, carboxymethyl cellulose, sodium alginate, and the like), a preservative (e.g., methyl paraben, propyl paraben, benzyl alcohol, and the like), an isotonizing agent (e.g., sodium chloride, mannitol, sorbitol, glucose, and the like), and the like, or as an oily injection in solution, suspension or emulsion in a vegetable oil such as olive oil, sesame oil, cottonseed oil or corn oil; propylene glycol and the like.

An oral preparation can be produced by formulating the compound of the present invention by a compression molding after addition of an excipient (e.g., lactose, sucrose, starch, and the like), a disintegrant (e.g., starch, calcium carbonate, and the like), a binder (e.g., starch, gum arabic, carboxymethyl cellulose, polyvinylpyrrolidone, hydroxypropyl cellulose, and the like), a lubricant (e.g., talc, magnesium stearate, polyethylene glycol 6000, and the like) and other additives, and, where necessary, coating the formulated product for the purpose of taste masking, enteric dissolution or sustained release by a commonly known method. Coating agents for this purpose include, for example, hydroxypropylmethyl cellulose, ethyl cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose, polyoxyethylene glycol, Tween 80, Prulonic F68, cellulose acetate phthalate, hydroxypropylmethyl cellulose phthalate, hydroxymethyl cellulose acetate succinate, Eudragit (produced by Rohm Company, Germany; methacrylic acid/acrylic acid copolymer), dyes (e.g., iron oxide, titanium dioxide), and the like. For an enteric preparation, an intermediate phase may be provided between the enteric phase and the drug-containing phase for the purpose of separation of the two phases by a commonly known method.

An external preparation can be produced by converting the compound of the present invention as a solid, semi-solid or liquid composition by a commonly known method. Such a solid composition is produced by, for example, powdering the compound of the present invention as such or in mixture with an excipient (e.g., glycol, mannitol, starch, microcrystalline cellulose, and the like), a thickening agent (e.g., natural rubber, cellulose derivative, acrylic acid polymer, and the like) and other additives. Such a liquid composition is produced by preparing the compound of the present invention as an oily or aqueous suspension in almost the same manner as the injection. The semi-solid composition is preferably an aqueous or oily gel, or an ointment. All these compositions may contain pH regulators (e.g., carbonic acid, phosphoric acid, citric acid, hydrochloric acid, sodium hydroxide, and the like), preservatives (e.g., paraoxybenzoic acid esters, chlorobutanol, benzalkonium chloride, and the like) and other additives.

A suppository is produced by preparing the compound of the present invention as an oily or aqueous solid, semi-solid or liquid composition by a commonly known method. Useful oily bases for such compositions include glycerides of higher fatty acids (e.g., cacao fat, witepsols (produced by Dynamite Nobel Company, Germany) and the like; medium fatty acids (e.g., MIGLIOL, produced by Dynamite Nobel Company, Germany); and vegetable oils (e.g., sesame oil, soybean oil, cottonseed oil, and the like). Aqueous bases include, for example, polyethylene glycols and propylene glycol. Bases for aqueous gels include, for example, natural rubbers, cellulose derivatives, vinyl polymers and acrylic acid polymers.

A compound of the formula:

wherein R11 and R13 each is a C1-4alkyl, R14 is a hydrogen atom or a C1-4alkoxy and m is an integer of 1 to 4 (hereinafter briefly referred to as Compound (V)) or a salt thereof also has an excellent GnRH antagonizing activity, especially a strong antagonistic activity though the compound falls outside the scope of Compound (I).

Preferable examples of Compound (V) are N-(4-(5-(((2-methoxyethyl)methylamino)methyl)-1-(2,6-difluorobenzyl)-1,2,3,4-tetrahydro-2,4-dioxo-3-(4-methoxyphenyl)thieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea or N-(4-(1-(2,6-difluorobenzyl)-5-((dimethylamino)methyl)-3-(4-methoxyphenyl)-2,4-dioxo-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea and a salt thereof.

Compound (V) can be produced by the method described in JP-A-9-169768 (WO 96/24597) and JP-A-2001-278884 (WO 00/56739) and its analogous method.

Compound (V) may be a hydrate or a non-hydrate. The hydrate is exemplified by monohydrate, sesquihydrate and dihydrate.

Compound (V) can be used as a prodrug. The prodrug means a compound which is converted to Compound (V) by a reaction due to an enzyme, a gastric acid, or the like under physiological conditions in the living body, that is, a compound which is converted to Compound (V) with oxidation, reduction, hydrolysis, or the like according to an enzyme, or a compound which is converted to Compound (V) with gastric acid, etc. The prodrug for Compound (V) may for example be a compound obtained by subjecting an amino group in Compound (V) to an acylation, alkylation or phosphorylation (e.g., a compound obtained by subjecting an amino group in Compound (V) to an eicosanoylation, alanylation, pentylaminocarbonylation, (5-methyl-2-oxo-1,3-dioxolen-4-yl)methoxycarbonylation, tetrahydrofuranylation, pyrrolidylmethylation, pivaloyloxymethylation and tert-butylation, etc.). These compounds can be produced from the compound of the present invention.

A prodrug of Compound (V) may also be one which is converted into Compound (V) under a physiological condition, such as those described in “IYAKUHIN no KAIHATSU (Development of Pharmaceuticals)”, Vol. 7, Design of Molecules, p. 163-198, Published by HIROKAWA SHOTEN (1990).

Compound (V) may be labeled with an isotope (e.g., 3H, 14C, 35S) and the like.

And the compound represented by the formula:

wherein Ra is (1) a hydrogen atom, (2) an aryl group which may have 1 to 5 substituent(s) selected from the group consisting of (i) a halogen, (ii) a nitro, (iii) a cyano, (iv) an amino, (v) a carboxyl group which may be esterified or amidated, (vi) an alkylenedioxy, (vii) an alkyl, (viii) an alkoxy, (ix) an alkylthio, (x) an alkylsulfinyl and (xi) an alkylsulfonyl, (3) a cycloalkyl group which may have a substituent or (4) a heterocyclic group which may have a substituent; Rb is a nitrogen-containing heterocyclic group which may have a substituent; Rc is an amino group which may have a substituent; Rd is an aryl group which may have a substituent; p is an integer of 0 to 3; and q is an integer of 0 to 3 (hereinafter as abbreviated as Compound (A)) or a salt thereof, which contains a part of the Compound (I) of the present invention, has an excellent GnRH antagonizing activity, especially strong antagonistic activity similar to Compound (I).

The definitions of the substituents of Compound (A) are shown below.

Examples of “aryl” of “aryl group which may have 1 to 5 substituent(s) selected from the group consisting of (i) a halogen, (ii) a nitro, (iii) a cyano, (iv) an amino, (v) a carboxyl group which may be esterified or amidated, (vi) an alkylenedioxy, (vii) an alkyl, (viii) an alkoxy, (ix) an alkylthio, (x) an alkylsulfinyl and (xi) an alkylsulfonyl” represented by Ra are C6-14aryl such as phenyl, 1-naphthyl, 2-naphthyl, anthryl, phenanthryl, acenaphthylenyl, and the like.

Examples of “halogen” of “aryl group which may have 1 to 5 substituent(s) selected from the group consisting of (i) a halogen, (ii) a nitro, (iii) a cyano, (iv) an amino, (v) a carboxyl group which may be esterified or amidated, (vi) an alkylenedioxy, (vii) an alkyl, (viii) an alkoxy, (ix) an alkylthio, (x) an alkylsulfinyl and (xi) an alkylsulfonyl” represented by Ra are fluorine, chlorine, bromine and iodine.

Examples of “carboxyl group which may be esterified or amidated” of “aryl group which may have 1 to 5 substituent(s) selected from the group consisting of (i) a halogen, (ii) a nitro, (iii) a cyano, (iv) an amino, (v) a carboxyl group which may be esterified or amidated, (vi) an alkylenedioxy, (vii) an alkyl, (viii) an alkoxy, (ix) an alkylthio, (x) an alkylsulfinyl and (xi) an alkylsulfonyl” represented by Ra are carboxyl, a C1-6alkoxy-carbonyl (e.g., methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, sec-butoxycarbonyl, tert-butoxycarbonyl, pentyloxycarbonyl, hexyloxycarbonyl, and the like), a C3-6cycloalkyloxy-carbonyl (e.g., cyclopropyloxycarbonyl, cyclobutyloxycarbonyl, cyclopentyloxycarbonyl, cyclohexyloxycarbonyl, and the like), a C6-14aryloxy-carbonyl (e.g., phenoxycarbonyl, 1-naphthyloxycarbonyl, 2-naphthyloxycarbonyl, anthryloxycarbonyl, phenanthryloxycarbonyl, acenaphthylenyloxycarbonyl, and the like), a C7-10 aralkyloxy-carbonyl (e.g., benzyloxycarbonyl, and the like), carbamoyl, a N-mono-C1-6alkyl-carbamoyl (e.g., methylcarbamoyl, ethylcarbamoyl, propylcarbamoyl, isopropylcarbamoyl, butylcarbamoyl, isobutylcarbamoyl, sec-butylcarbamoyl, tert-butylcarbamoyl, pentylcarbamoyl, hexylcarbamoyl, and the like), a N-mono-C3-6cycloalkyl-carbamoyl (e.g., cyclopropylcarbamoyl, cyclobutylcarbamoyl, cyclopentylcarbamoyl, cyclohexylcarbamoyl, and the like), a N-mono-C6-14aryl-carbamoyl (e.g., phenylcarbamoyl, 1-naphthylcarbamoyl, 2-naphthylcarbamoyl, anthrylcarbamoyl, phenanthryloxycarbamoyl, acenaphthylenyloxycarbamoyl, and the like), N-mono-C7-10aralkyl-carbamoyl (e.g., benzylcarbamoyl, and the like), and the like.

Examples of “alkylenedioxy” of “aryl group which may have 1 to 5 substituent(s) selected from the group consisting of (i) a halogen, (ii) a nitro, (iii) a cyano, (iv) an amino, (v) a carboxyl group which may be esterified or amidated, (vi) an alkylenedioxy, (vii) an alkyl, (viii) an alkoxy, (ix) an alkylthio, (x) an alkylsulfinyl and (xi) an alkylsulfonyl” represented by Ra are a C1-6alkylenedioxy (e.g., —OCH2O—, —O(CH2)2O—, —O(CH2)3O—, —O(CH2)4O—, —O(CH2)4O—, —O(CH2)6O—).

Examples of “alkyl” of “aryl group which may have 1 to 5 substituent(s) selected from the group consisting of (i) a halogen, (ii) a nitro, (iii) a cyano, (iv) an amino, (v) a carboxyl group which may be esterified or amidated, (vi) an alkylenedioxy, (vii) an alkyl, (viii) an alkoxy, (i) an alkylthio, (x) an alkylsulfinyl and (xi) an alkylsulfonyl” represented by Ra are a C1-6alkyl (e.g., methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, and the like), and the like.

Examples of “alkoxy” of “aryl group which may have 1 to 5 substituent(s) selected from the group consisting of (i) a halogen, (ii) a nitro, (iii) a cyano, (iv) an amino, (v) a carboxyl group which may be esterified or amidated, (vi) an alkylenedioxy, (vii) an alkyl, (viii) an alkoxy, (ix) an alkylthio, (x) an alkylsulfinyl and (xi) an alkylsulfonyl” represented by Ra are a C1-6alkoxy (e.g., methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, hexyloxy, and the like), and the like.

Examples of “alkylthio” of “aryl group which may have 1 to 5 substituent(s) selected from the group consisting of (i) a halogen, (ii) a nitro, (iii) a cyano, (iv) an amino, (v) a carboxyl group which may be esterified or amidated, (vi) an alkylenedioxy, (vii) an alkyl, (viii) an alkoxy, (ix) an alkylthio, (x) an alkylsulfinyl and (xi) an alkylsulfonyl” represented by Ra are a C1-6alkylthio (e.g., methylthio, ethylthio, propylthio, isopropylthio, butylthio, isobutylthio, sec-butylthio, tert-butylthio, pentylthio, hexylthio, and the like), and the like.

Examples of “alkylsulfinyl” of “aryl group which may have 1 to 5 substituent(s) selected from the group consisting of (i) a halogen, (ii) a nitro, (iii) a cyano, (iv) an amino, (v) a carboxyl group which may be esterified or amidated, (vi) an alkylenedioxy, (vii) an alkyl, (viii) an alkoxy, (ix) an alkylthio, (x) an alkylsulfinyl and (xi) an alkylsulfonyl” represented by Ra are a C1-6alkylsulfinyl (e.g., methylsulfinyl, ethylsulfinyl, propylsulfinyl, isopropylsulfinyl, butylsulfinyl, isobutylsulfinyl, sec-butylsulfinyl, tert-butylsulfinyl, pentylsulfinyl, hexylsullfinyl, and the like), and the like.

Examples of “alkylsulfonyl” of “aryl group which may have 1 to 5 substituent(s) selected from the group consisting of (i) a halogen, (ii) a nitro, (iii) a cyano, (iv) an amino, (v) a carboxyl group which may be esterified or amidated, (vi) an alkylenedioxy, (vii) an alkyl, (viii) an alkoxy, (ix) an alkylthio, (x) an alkylsulfinyl and (xi) an alkylsulfonyl” represented by Ra are a C1-6alkylsulfonyl (e.g., methylsulfonyl, ethylsulfonyl, propylsulfonyl, isopropylsulfonyl, butylsulfonyl, isobutylsulfonyl, sec-butylsulfonyl, tert-butylsulfonyl, pentylsulfonyl, hexylsulfonyl, and the like), and the like.

Examples of “cycloalkyl group” of “cycloalkyl group which may have a substituent” represented by Ra are a C3-6cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like), and the like.

Examples of “substituent” of “nitrogen-containing heterocyclic group which may have a substituent” represented by Rb are same number and same kind as “substituent” of “cycloalkyl group which may have a substituent” and “heterocyclic group which may have a substituent” represented by Ra.

Example of “amino group which may have a substituent” represented by Rc is a group of the formula: —NReRf wherein Re is (1) a hydrogen atom, (2) a C1-6alkyl which may have a substituent, (3) a C3-6cycloalkyl which may have a substituent, (4) a C6-14aryl which may have a substituent, (5) a C7-20aralkyl which may have a substituent, (6) a carbamoyl which may have a substituent or (7) a heterocyclic group; and Rf is a hydrogen atom or a C1-6alkyl whish may have a substituent.

Examples of “C1-16alkyl” of “C1-6alkyl which may have a substituent” represented by Re and Rf are methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, and the like.

Examples of “C3-6cycloalkyl” of “C3-6cycloalkyl which may have a substituent” represented by Re are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

Examples of the substituent of “C3-6cycloalkyl which may have a substituent” represented by Re are the same as the substituent of “C1-6alkyl which may have a substituent” represented by Re and Rf above. The number of substituents is 1 to 6, preferably 1 to 3, and the substitution position may be any place on which the substitution is possible.

Examples of C6-14aryl of “C6-14aryl which may have a substituent” represented by Re are phenyl, naphthyl, anthracenyl, and the like.

Examples of the substituent of “C6-14aryl which may have a substituent” are the same as the substituent of “C1-6alkyl which may have a substituent” represented by Re and Rf above except for oxo and epoxy. The number of substituents is 1 to 6, preferably 1 to 3, and the substitution position may be any place on which the substitution is possible.

Examples of C7-20aralkyl of “C7-20aralkyl which may have a substituent” represented by Re are benzyl, phenethyl, phenylpropyl, benzhydoryl, trityl and the like.

Examples of the substituent of “C7-20aralkyl which may have a substituent” are the same as the substituent of “C1-6alkyl which may have a substituent” represented by Re and Rf above. The number of substituent is 1 to 6, preferably 1 to 3, and the substitution position may be any place on which the substitution is possible.

Examples of the substituent of “carbamoyl which may have a substituent” represented by Re are (1) a C1-6alkyl which may have a substituent, (2) a C3-6cycloalkyl which may have a substituent, (3) a C6-14aryl which may have a substituent, (4) a C7-20aralkyl which may have a substituent, (5) a hydroxy, (6) a C1-6alkoxy which may have a substituent, (7) a C1-6alkoxy-carbonyl which may have a substituent, and the like. The number of the substituent may be 1 or 2.

Examples of “C1-16alkyl which may have a substituent” as a substituent of “carbamoyl which may have a substituent” represented by Re are the same as “C1-6alkyl which may have a substituent” represented by Re and Rf above.

Examples of “C3-6cycloalkyl which may have a substituent”, “C6-14aryl which may have a substituent” and “C7-20aralkyl which may have a substituent” as a substituent of “carbamoyl which may have a substituent” represented by Re are the same as “C3-6cycloalkyl which may have a substituent”, “C6-14aryl which may have a substituent” and “C7-20aralkyl which may have a substituent” represented by Re above.

Examples of the C1-6alkoxy of “C1-6alkoxy which may have a substituent” as a substituent of “carbamoyl which may have a substituent” represented by Re are methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, hexyloxy, and the like. Examples of the substituent of “C1-6alkoxy which may have a substituent” are the same as the substituent of “C1-6alkyl which may have a substituent” represented by Re above. The number of substituents is 1 to 6, preferably 1 to 3, and the substitution position may be any place on which the substitution is possible.

Example of “C1-6alkoxy-carbonyl which may have a substituent” as a substituent of “carbamoyl which may have a substituent” represented by Re is a group comprising combining “C1-6alkoxy which may have a substituent” as a substituent of “carbamoyl which may have a substituent” represented by Re above with carbonyl.

Examples of the aryl of “aryl which may have a substituent” represented by Rd are phenyl, naphthyl, anthracenyl, and the like.

Examples of the substituent of “aryl which may have a substituent” represented by Rd are (1) a C6-14aryl (e.g., phenyl, naphthyl, and the like) which may have 1 to 4 substituent(s) selected from the group consisting of (i) a hydroxy, (ii) an amino, (iii) a mono- or di-C1-6alkylamino (e.g., methylamino, ethylamino, propylamino, dimethylamino, diethylamino, and the like), (iv) a C1-6alkoxy (e.g., methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, and the like), and (v) a halogen (e.g., fluorine, chlorine, bromine and iodine), (2) a hydroxy, (3) a carboxy, (4) a nitro, (5) a C1-6alkoxy (e.g., methoxy, ethoxy, propoxy, isopropoxy, butoxy, pentyloxy, hexyloxy, and the like), (6) a C1-6alkyl-carbonyloxy (e.g., acetoxy, propionyloxy, butyryloxy, isobutyryloxy, valeryloxy, isovaleryloxy, pivaloyloxy, pentylcarbonyloxy, hexylcarbonyloxy, and the like), (7) a C1-6alkylthio (e.g., methylthio, ethylthio, propylthio, isopropylthio, butylthio, isobutylthio, sec-butylthio, tert-butylthio, pentylthio, hexylthio, and the like), (8) a C1-6alkylsulfinyl (e.g., methylsulfinyl, ethylsulfinyl, propylsulfinyl, isopropylsulfinyl, butylsulfinyl, isobutylsulfinyl, sec-butylsulfinyl, tert-butylsulfinyl, pentylsulfinyl, hexylsulfinyl, and the like), (9) a C1-6alkylsulfonyl (e.g., methylsulfonyl, ethylsulfonyl, propylsulfonyl, isopropylsulfonyl, butylsulfonyl, isobutylsulfonyl, sec-butylsulfonyl, tert-butylsulfonyl, pentylsulfonyl, hexylsulfonyl, and the like), (10) a halogen (e.g., fluorine, chlorine, bromine and iodine), (11) a group of the formula: —NRgRh wherein Rg is (a) a hydrogen atom, (b) a C1-6alkyl which may have a substituent, (c) a C3-6cycloalkyl which may have a substituent, (d) a C6-14aryl which may have a substituent, (e) a C7-20aralkyl which may have a substituent, (a carbamoyl which may have 1 or 2 substituent(s) selected from the group consisting of (i) a C3-6cycloalkyl which may have a substituent, (ii) a C6-14aryl which may have a substituent, (iii) a C7-20 aralkyl which may have a substituent, (iv) hydroxy, (v) a C1-6alkoxy which may have a substituent and (vi) a C1-6alkoxy-carbonyl which may have a substituent, (g) a heterocyclic group; and Rh is a hydrogen atom or a C1-6alkyl which may have a substituent, (12) a 5-membered cyclic group containing 1 to 4 heteroatom(s) selected from an oxygen atom, a sulfur atom, a nitrogen atom, and the like in addition to carbon atom (e.g., 2-thienyl, 3-thienyl, 2-furyl, 3-furyl, 2-pyrrolyl, 3-pyrrolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 3-pyrazolyl, 4-pyrazolyl, 5-pyrazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 3-isothiazolyl, 4-isothiazolyl, 5-isothiazolyl, 3-(1,2,4-oxadiazolyl), 5-(1,2,4-oxadiazolyl), 1,3,4-oxadiazolyl, 3-(1,2,4-thiadiazolyl), 5-(1,2,4-thiadiazolyl), 1,3,4-thiadiazolyl, 4-(1,2,3-thiadiazolyl), 5-(1,2,3-thiadiazolyl), 1,2,5-thiadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1H-tetrazolyl, 2H-tetrazolyl, oxoimidazinyl, dioxotriazinyl, pyrrolidinyl, and the like), (13) a 6-membered cyclic group containing 1 to 4 heteroatom(s) selected from an oxygen atom, a sulfur atom, a nitrogen atom, and the like in addition to carbon atom (e.g., 2-pyridyl, 3-pyridyl, 4-pyridyl, N-oxido-2-pyridyl, N-oxido-3-pyridyl, N-oxido-4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, N-oxido-2-pyrimidinyl, N-oxido-4-pyrimidinyl, N-oxido-5-pyrimidinyl, 2-thiomorpholinyl, 3-thiomorpholinyl, 2-morpholinyl, 3-morpholinyl, piperidinyl, pyranyl, thiopyranyl, 1,4-oxazinyl, 1,4-thiazinyl, 1,3-thiazinyl, 2-piperazinyl, 3-piperazinyl, triazinyl, oxotriazinyl, 3-pyridazinyl, 4-pyridazinyl, pyrazinyl, N-oxido-3-pyridazinyl, N-oxido-4-pyridazinyl, and the like), (14) a bicyclic or tricyclic condensed cyclic group which contains 1 to 4 heteroatom(s) selected from an oxygen atom, a sulfur atom, a nitrogen atom and the like in addition to a carbon atom (e.g., benzofuryl, benzothiazolyl, benzoxazolyl, tetrazolo[1,5-b]pyridazinyl, triazolo[4,5-b]pyridazinyl, benzoimidazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, indolizinyl, quinolizinyl, 1,8-naphthylidinyl, purinyl, pteridinyl, dibenzofuranyl, carbazolyl, acridinyl, phenanthrydinyl, chromanyl, benzoxazinyl, phenazinyl, phenothiazinyl, phenoxazinyl, and the like), (15) a C1-6alkoxycarbonyl (e.g., methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, sec-butoxycarbonyl, tert-butoxycarbonyl, pentyloxycarbonyl, hexyloxycarbonyl, and the like), (16) a carbamoyl, (17) a N-mono-C1-6alkylcarbamoyl (e.g., N-methylcarbamoyl, N-ethylcarbamoyl, N-propylcarbamoyl, N-isopropylcarbamoyl, and the like), (18) a N,N-di-C1-6alkylcarbamoyl (e.g., N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N,N-dipropylcarbamoyl, and the like), and the like. The number of substituent is 1 to 6, preferably 1 to 3, and the substitution position may be any place on which the substitution is possible.

The definitions of R8 and Rf utilized in the group of the formula: —NRgRh wherein Rg and Rh have the same meanings defined above as a substituent of “aryl which may have a substituent” represented by Rd are shown below.

Examples of “C1-6alkyl which may have a substituent” represented by Rg and Rh are the same as “C1-6alkyl which may have a substituent” represented by Re and Rf described above.

Examples of “C3-6cycloalkyl which may have a substituent”, “C6-14aryl which may have a substituent”, “C7-20aralkyl which may have a substituent” and “heterocyclic group” represented by Rg are the same as “C3-6cycloalkyl which may have a substituent”, “C6-14aryl which may have a substituent”, “C7-20aralkyl which may have a substituent” and “heterocyclic group” represented by Re described above.

Examples of “C3-6cycloalkyl which may have a substituent”, “C6-14aryl which may have a substituent” and “C7-20aralkyl which may have a substituent” of “carbamoyl which may have 1 or 2 substituent(s) selected from the group consisting of (i) a C3-6cycloalkyl which may have a substituent, (ii) a C6-14aryl which may have a substituent, (iii) a C7-20aralkyl which may have a substituent, (iv) a hydroxy, (v) a C1-6alkoxy which may have a substituent and (vi) a C1-6alkoxy-carbonyl which may have a substituent” represented by Rg are the same as “C3-6cycloalkyl which may have a substituent”, “C6-14aryl which may have a substituent” and “C7-20aralkyl which may have a substituent” represented by Re described above.

Examples of “C1-6alkoxy which may have a substituent” of “carbamoyl which may have 1 or 2 substituent(s) selected from the group consisting of (i) a C3-6cycloalkyl which may have a substituent, (ii) a C6-14aryl which may have a substituent, (iii) a C7-20aralkyl which may have a substituent, (iv) a hydroxy, (v) a C1-6alkoxy which may have a substituent and (vi) a C1-6alkoxy-carbonyl which may have a substituent” represented by Rg are methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, hexyloxy, and the like. Examples of the substituent of said “C1-6alkoxy which may have a substituent” are the same as the substituent of “C1-6alkyl which may have a substituent” represented by Re described above. The number of substituents is 1 to 6, preferably 1 to 3, and the substitution position may be any place on which the substitution is possible.

Example of “C1-6alkoxy-carbonyl which may have a substituent” of “carbamoyl which may have 1 or 2 substituent(s) selected from the group consisting of (i) a C3-6cycloalkyl which may have a substituent, (ii) a C6-14aryl which may have a substituent, (iii) a C7-20aralkyl which may have a substituent, (iv) a hydroxy, (v) a C1-6alkoxy which may have a substituent and (vi) a C1-6alkoxy-carbonyl which may have a substituent” represented by Rg is a group comprising combining “C1-6alkoxy which may have a substituent” as a substituent of “carbamoyl which may have a substituent” represented by Rg above with carbonyl.

Preferable examples of Ra are an aryl group which may have 1 to 5 substituent(s) selected from the group consisting of (i) a halogen, (ii) a nitro, (iii) a cyano, (iv) an amino, (v) a carboxyl group which may be esterified or amidated, (vi) an alkylenedioxy, (vii) an alkyl, (viii) an alkoxy, (ix) an alkylthio, (x) an alkylsulfinyl and (xi) an alkylsulfonyl. Among them, a phenyl which is mono- or di-substituted by a halogen, especially 2,6-difluorophenyl, are preferable.

Preferable example of Rc is a group represented by the formula: —NRe′Rf wherein Re′ is (1) a C1-6alkyl which may have a substituent or (2) a C7-20aralkyl; Rf′ is a C1-6alkyl. Among them, a group of the formula: —N(Me)Re″ wherein Re″ is a C1-6alkyl which is substituted by a C1-6alkoxy; or benzyl is preferable.

Preferable example of Rd is phenyl which may have a substituent. Among them, phenyl group which is substituted at its 4-position by a group represented by the formula —NRgRh: wherein each Rg and Rh has the meaning defined above, is preferable, and especially, phenyl group which is substituted at its 4-position by a group represented by the formula —NHRg′ wherein Rg′ is carbamoyl which may have 1 or 2 substituent(s) selected from the group consisting of (i) a C3-6cycloalkyl which may have a substituent, (ii) a C6-14aryl which may have a substituent, (iii) a C7-20aralkyl which may have a substituent, (iv) hydroxy, (v) a C1-6alkoxy which may have a substituent and (vi) a C1-6alkoxy-carbonylamino is preferable. Especially, a phenyl group substituted at its 4-position by a C1-6alkoxyamino-carbonylamino (e.g. 4-methoxyaminocarbonylaminophenyl, 4-ethoxyaminocarbonylaminophenyl, and the like) is more preferable.

Compound (A) can be produced by the method described in JP-A-9-169768 (WO 96/24597) and JP-A-2001-278884 (WO 00/56739) and its analogous method.

Compound (A) may be a hydrate or a non-hydrate. The hydrate is exemplified by monohydrate, sesquihydrate and dihydrate.

When Compound (A) is obtained as a mixture of optically active configurations, it can be resolved into the (R)- and (S)-forms by the conventional optical resolution techniques.

Compound (A) can be used as a prodrug. The prodrug of Compound (A) or a salt thereof means a compound which is converted to Compound (A) of the present invention under physiological conditions or with a reaction due to an enzyme, a gastric acid, and the like in the living body, that is, a compound which is converted to Compound (A) of the present invention with oxidation, reduction, hydrolysis, and the like with an enzyme; a compound which is converted to Compound (A) of the present invention with gastric acid, etc. The prodrug for Compound (A) may for example be a compound obtained by subjecting an amino group in Compound (A) to an acylation, alkylation or phosphorylation (e.g., a compound obtained by subjecting an amino group in Compound (A) or to an eicosanoylation, alanylation, pentylaminocarbonylation, (5-methyl-2-oxo-1,3-dioxolen-4-yl)methoxycarbonylation, tetrahydrofuranylation, pyrrolidylmethylation, pivaloyloxymethylation and tert-butylation, etc.); a compound obtained by subjecting a hydroxy group in Compound (A) to an acylation, alkylation, phosphorylation or boration (e.g., a compound obtained by subjecting an hydroxy in Compound (A) to an acetylation, palmitoylation, propanoylation, pivaloylation, succinylation, fumarylation, alanylation, dimethylaminomethylcarbonylation, etc.); a compound obtained by subjecting a carboxy group in Compound (A) to an esterification or amidation (e.g., a compound obtained by subjecting a carboxyl group in compound (A) to an ethylesterification, phenylesterification, carboxymethylesterification, dimethylaminomethylesterification, pivaloyloxymethylesterification, ethoxycarbonyloxyethylesterification, phthalidylesterification, (5-methyl-2-oxo-1,3-dioxolen-4-yl)methylesterification, cyclohexyloxycarbonylethylesterification and methylamidation, etc.) and the like. Any of these compounds can be produced from the compound of the present invention by a method known per se.

A prodrug of Compound (A) may also be one which is converted into Compound (A) under a physiological condition, such as those described in “IYAKUHIN no KAIHATSU (Development of Pharmaceuticals)”, Vol. 7, Design of Molecules, p. 163-198, Published by HIROKAWA SHOTEN (1990).

Compound (A) may be labeled with an isotope (e.g., 3H, 14C, 35S) and the like.

The present invention is hereinafter described in more detail by means of, but is not limited to, the following Reference Examples, Examples, Preparation Examples and Experimental Examples.

1H-NMR spectra are determined with tetramethylsilane as the internal standard, using the Varian GEMINI 200 (200 MHz) spectrometer, the JEOL LAMBDA 300 (300 MHz) spectrometer or the Bruker AM500 (500 MHz) spectrometer; all δ values are shown in ppm. Unless otherwise specifically indicated, “%” is by weight. Yield indicates mol/mol %. The other symbols used herein have the following definitions:

s: singlet

d: doublet

t: triplet

dt: double triplet

m: multiplet

br: broad

AIBN: 2,2-azobisisobutyronitrile

DMF: N,N-dimethylformamide

NBS: N-bromosuccinimide

TFA: trifluoroacetic acid

THF: tetrahydrofuran

Me: methyl

Et: ethyl

Ph: phenyl

TBS: tert-butyl dimethyl silyl

Ms: methanesulfonyl

The term “room temperature” indicates the range from about 15 to 25° C., but is not to be construed as strictly limitative. Each of lactose, corn starch, D-mannitol, low substituted hydroxypropylcellulose, talc, hydroxypropylcellulose, hydroxypropylmethylcellulose 2910, titanium oxide and light silicic acid anhydride used in the following Preparation is suited for standard of Pharmacopoeia, Fourteenth Edition.

To a solution of ethyl 4-(N-benzyl-N-methylaminomethyl)-2-[N-(2,6-difluorobenzyl)-N-ethoxycarbonylamino]-5-[4-(3-methoxyureido)phenyl]thiophene-3-carboxylate (3.64 g, 5.47 mmol) in ethanol (100 ml) were added 1N hydrochloric acid (8 ml) and 10% palladium-carbon (50% wet, 1.82 g). The mixture was stirred vigorously under hydrogen atmosphere for 6 hours. The catalyst was removed, and the filtrate was neutralized with 1N sodium hydroxide solution. The solvent was distilled off, and the residue was distributed between ethyl acetate and water. The organic layer was washed with saturated brine and dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure. The residue was subjected to an NH-silica gel (Fuji Silysia Chemical) chromatography to give the title compound (2.43 g, 77%) as yellow powders.

To a solution of the compound obtained in Reference Example 2 (2.34 g, 3.5 mmol) in ethanol (40 ml) was added 2N sodium hydroxide solution (8.75 ml), and the mixture was stirred at 50-60° C. for 14 hours. The reaction mixture was cooled to room temperature and neutralized with 1N hydrochloric acid. The solvent was distilled off and the obtained residue was distributed between ethyl acetate and water. The organic layer was washed with saturated brine and dried over magnesium sulfate. The solvent was distilled off under reduced pressure to give the title compound (2.06 g, 92%) as pale yellow powders.

A solution of 2N hydrochloride in diethyl ether (21 ml) and 10% palladium-carbon (50% wet, 3.73 g) were added to a solution of ethyl 2-[N-(2,6-difluorobenzyl)-N-ethoxycarbonylamino]-4-[N-(2-methoxyethyl)-N-methylaminomethyl]-5-(4-nitrophenyl)thiophene-3-carboxylate (12.43 g) (JP-A-2001-278884, WO 00/56739) in ethanol (315 ml). The mixture was stirred vigorously under hydrogen atmosphere for 1 hour. The catalyst was removed, and the filtrate was neutralized with sodium hydrogen carbonate solution. The solvent was distilled off, and the residue was distributed between ethyl acetate and water. The organic layer was washed with saturated brine and dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure. The residue was subjected to an NH-silica gel (Fuji Silysia Chemical) chromatography to give the title compound (11.44 g) as an oil.

N-ethyldiisopropylamine (3.06 ml) was added to a solution of the compound obtained in Reference Example 5 (4.89 g) in dichloromethane (113 ml) under ice cooling, and the mixture was stirred. N,N′-carbonyldiimidazole (2.82 g) was added to the mixture under ice cooling. The reaction mixture was warmed to room temperature and stirred for 67 hours. The reaction mixture was cooled under ice cooling, and O-methylhydroxyamine hydrochloride (7.26 g) and N-ethyldiisopropylamine (15.6 ml) were added thereto. The reaction mixture was warmed to room temperature and stirred at room temperature for 19 hours. The reaction mixture was distributed between chloroform and saturated aqueous solution of sodium hydrogen carbonate, and extracted with chloroform. The combined extract was washed with brine and dried over magnesium sulfate. The solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography to give the title compound (4.89 g) as a pale yellow caramelized product.

An aqueous solution of 2N sodium hydroxide (18.9 ml) was added to a solution of the compound obtained in Reference Example 6 (4.81 g) in ethanol (114 ml), and the mixture was stirred at 60° C. for 5 hours. The reaction mixture was warmed to room temperature and combined with 1N hydrochloric acid (37.8 ml). The solvent was distilled off. The residue was dissolved in ethanol and toluene, and the solvent was distilled off. The residue was combined with anhydrous ethanol (30 ml), and the inorganic products were filtered off. The filtrate was concentrated to dryness. The obtained residue was fined by anhydrous ether, collected by filtration and dried to give the title compound (4.43 g).

Potassium carbonate (4.15 g) and methyl iodide (2.80 ml) were added to a solution of 6-methyl-2-nitro-3-hydroxypyridine (4.63 g) in DMF (120 ml), and the mixture was stirred at room temperature for 14 hours. The reaction mixture was distributed between ethylacetate and water. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was recrystallized from ethyl acetate/hexane to give the title compound (3.94 g) as needle crystals.

To a solution of 3-methoxy-6-methyl-2-nitropyridine (3.85 g) in ethanol (91.6 ml) was added 10% palladium-carbon (50% wet, 0.96 g), and the mixture was stirred under hydrogen atmosphere for 2 hours. The catalyst was filtered off, and the filtrate was concentrated to dryness. The residue was recrystallized from ethyl acetate/hexane to give the title compound (2.89 g).

The similar reaction as described in Reference Example 9 by using 2-nitro-3-hydroxypyridine (7.0 g), potassium carbonate (6.91 g) and methyl iodide (4.67 ml) gave the title compound (7.5 g) as crystalline powders.

1H-NMR (CDCl3) δ: 3.99 (3H, s), 7.54-7.56 (2H, m), 8.09-8.12 (1H, m).

IR (KBr): 1601, 1537, 1530, 1422, 1364, 1275 cm−1.

Reference Example 13Production of 2-amino-3-methoxypyridine

The similar reaction as described in Reference Example 10 by using 3-methoxy-2-nitropyridine (7.5 g) gave the title compound (5.42 g) as crystalline powders.

To a solution of the compound obtained in Example 1 (1.59 g, 2.38 mmol) in ethanol (40 ml) were added 1N hydrochloric acid (7 ml) and 10% palladium-carbon (50% wet, 0.63 g), and the mixture was stirred vigorously under hydrogen atmosphere for 20 hours. The catalyst was removed, and the filtrate was neutralized with 1N sodium hydroxide solution. The solvent was distilled off. The residue was distributed between ethyl acetate and water, and the organic layer was washed with saturated brine and dried over anhydrous magnesium sulfate. The solvent was distilled off, and the obtained powders were washed with diethyl ether to give the title compound (980 mg, 71%) as pale yellow powders.

2-Methylamino)ethanol (0.14 g, 1.903 mmol) was dissolved in THF (10 ml), and triethylamine (0.58 ml, 4.15 mmol) and methanesulfonyl chloride (0.27 ml, 3.46 mmol) were added thereto. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was combined with an aqueous solution of sodium hydrogen carbonate and extracted with ethyl acetate. The water layer was salted out and extracted with ethyl acetate. The combined organic layer was dried over magnesium sulfate and concentrated under reduced pressure to give a mesylate. A solution of the obtained mesylate, the compound obtained in Reference Example 14 (200 mg, 0.346 mmol), N,N-diisopropylethylamine (0.12 ml, 0.692 mmol) and potassium iodide (230 mg, 1.38 mmol) in DMF (8 ml) was stirred at 50-60° C. for 16 hours. The reaction mixture was combined with an aqueous solution of sodium hydrogen carbonate and extracted with ethyl acetate. The organic layer was washed with brine, dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent:ethyl acetate/methanol=80/1) and recrystallized from dichloromethane/methanol/diethyl ether to give the title compound (115 mg, 47%) as pale yellow crystals.

The similar reaction as described in Example 2 by using the compound obtained in Reference Example 14 (200 mg, 0.346 mmol) and 1-(2-hydroxyethyl)-2-pyrrolidone (0.25 g, 1.903 mmol) gave the title compound (97 mg, 41%) as colorless crystals.

Production of N-(4-(5-((benzyl(methyl)amino)methyl)-1-(2,6-difuorobenzyl)-2,4-dioxo-3-(2-(1H-1,2,3-triazol-1-yl)ethyl)-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea (2)

The similar reaction as described in Example 2 by using the compound obtained in Example 5 (450 mg, 0.708 mmol) gave a mesylate. A solution of the obtained mesylate, 1,2,3-triazole (148 mg, 2.12 mmol and potassium carbonate (294 mg, 2.12 mmol) in DMF (8 ml) was stirred at room temperature for 18 hours, and at 50-60° C. for 3 hours. The reaction mixture was combined with an aqueous solution of sodium hydrogen carbonate and extracted with ethyl acetate. The organic layer was washed with brine, dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent:ethyl acetate) to give the title compound 2-yl form (243 mg, 50%) as a white amorphous compound. On the other hand, the residue was recrystallized from dichloromethane/methanol/diethyl ether to give the title compound 1-yl form (177 mg, 36%) as colorless crystals.

Production of N-(4-(5-((benzyl(methyl)amino)methyl)-1-(2,6-difluorobenzyl)-2,4-dioxo-3-(2-(1H-tetrazol-1-yl)ethyl)-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea (2)

The similar reaction as described in Example 6 by using the compound obtained in Reference Example 5 (636 mg, 1 mmol) and tetrazole (210 mg, 3 mmol) gave the title compound 2-yl form (234 mg, 34%) as a white amorphous compound. On the other hand, the residue was powdered by diethylether to give the title compound, 1-yl form, (34 mg, 5%) as pale yellow crystals.

The similar reaction as described in Example 5 by using 4-(N-benzyl-N-methylaminomethyl)-2-[N-(2,6-difluorobenzyl)-N-ethoxycarbonylamino]-5-[4-(3-methoxyureido)phenyl]thiophen-3-carboxylic acid (1.21 g, 2 mmol) and 2-aminoethanol (0.18 ml, 3 mmol) to give the title compound (302 mg, 25%) as a pale yellow amorphous compound.

Production of N-(4-(1-(2,6-difluorobenzyl)-5-(((2-methoxyethyl)(methyl)amino)methyl)-2,4-dioxo-3-(2-(1H-tetrazol-1-yl)ethyl)-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea (2)

The similar reaction as described in Example 6 by using the compound obtained in Example 8 (250 mg, 0.414 mmol) and tetrazole (145 mg, 2.07 mmol) gave the title compound 2-yl form (66 mg, 24%) and 1-yl form (27 mg, 10%) as white amorphous compounds.

The similar reaction as described in Reference Example 14 by using the compound obtained in Example 6 (1-yl form) (200 mg, 0.291 mmol) gave de-benzyl-form (0.11 g, 63%) as colorless powders. The similar reaction as described in Example 4 by using the de-benzyl-form (0.11 g, 0.184 mmol) and 2-(chloromethyl)methylether (0.10 ml, 1.104 mmol) gave the title compound (35 mg, 29%) as colorless crystals.

The similar reaction as described in Reference Example 14 by using 5-(N-benzyl-N-methylaminomethyl)-1-(2,6-difluorobenzyl)-6-[4-(3-methoxyureido)phenyl]-3-phenylthieno[2,3-d]pyrimidin-2,4(1H, 3H)-dione (6.68 g, 10 mmol) gave the title compound (5.52 g, 96%) as white powders.

Production of N-(4-(1-(2,6-difluorobenzyl)-5-((methyl(2-(1H-1,2,3-triazol-1-yl)ethyl)amino)methyl)-2,4-dioxo-3-phenyl-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea (2)

Potassium carbonate (1.33 g, 10 mmol) was added to a solution of 1,2,3-triazole (0.46 g, 6.67 mmol) and 1-bromo-2-chloroethane (0.83 ml, 10 mmol) in DMF (5 ml), and the mixture was stirred at room temperature for 1 hour, and 50-60° C. for 3 hours. The reaction mixture was combined with saturated brine and extracted twice with ethyl acetate. The combined organic layer was dried over magnesium sulfate and concentrated to give a halide.

The compound obtained in Reference Example 15 (700 mg, 1.21 mmol) was dissolved in DMF (12 ml), and N,N-diisopropylethylamine (1.16 ml, 6.67 mmol) and the halide obtained above were added thereto. The mixture was stirred at 50-60° C. for 16 hours, combined with an aqueous solution of sodium hydrogen carbonate and extracted with ethyl acetate. The organic layer was washed with brine, dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by NH-silica gel column chromatography (eluent:ethyl acetate) and recrystallized from dichloromethane/methanol/diethyl ether to give the title compound 2-yl form (140 mg, 17%) and 1-yl form (332 mg, 41%) as colorless crystals.

The similar reaction as described in Example 2 by using the compound obtained in Reference Example 15 (350 mg, 0.606 mmol) and 2-(2-hydroxyethyl)pyridine (0.45 g, 3.64 mmol) gave the title compound (233 mg, 56%) as colorless crystals.

The similar reaction as described in Example 2 by using the compound obtained in Reference Example 15 (350 mg, 0.606 mmol) and 4-(2-hydroxyethyl)pyridine hydrochloride (0.58 g, 3.64 mmol) gave the title compound (166 mg, 40%) as colorless crystals.

The similar reaction as described in Example 4 by using the compound obtained in Reference Example 15 (350 mg, 0.606 mmol) and 2-chloromethylpyridine hydrochloride (149 mg, 0.908 mmol) gave the title compound (297 mg, 73%) as colorless crystals.

The similar reaction as described in Example 5 by using 4-(N-benzyl-N-methylaminomethyl)-2-[N-(2,6-difluorobenzyl)-N-ethoxycarbonylamino]-5-[4-(3-methoxyureido)phenyl]thiophene-3-carboxylic acid (2.87 g, 4.49 mmol) and 4-fluoroaniline (0.64 ml, 6.735 mmol) gave the title compound (2.71 g, 88%) as pale yellow powders.

The similar reaction as described in Example 4 by using the compound obtained in Reference Example 17 (150 mg, 0.25 mmol) and 2-chloromethylpyridine hydrochloride (85 mg, 0.52 mmol) gave the title compound (105 mg, 61%) as colorless crystals.

The similar reaction as described in Example 2 by using the compound obtained in Reference Example 17 (150 mg, 0.25 mmol) and 2-(2-hydroxyethyl)pyridine (0.19 g, 1.512 mmol) gave the title compound (100 mg, 57%) as colorless crystals.

The similar reaction as described in Example 2 by using the compound obtained in Reference Example 14 (150 mg, 0.259 mmol) and 2-(2-hydroxyethyl)pyridine (0.18 g, 1.425 mmol) gave the title compound (83 mg, 47%) as pale yellow crystals.

The similar reaction as described in Example 4 by using the compound obtained in Reference Example 15 (150 mg, 0.26 mmol) and 3-chloromethylpyridine hydrochloride (85 mg, 0.52 mmol) gave the title compound (117 mg, 67%) as colorless crystals.

The similar reaction as described in Example 4 by using the compound obtained in Reference Example 15 (150 mg, 0.26 mmol) and 6-bromomethyl-2-pyridinemethanol (105 mg, 0.52 mmol) gave the title compound (115 mg, 63%) as colorless crystals.

The similar reaction as described in Example 4 by using the compound obtained in Reference Example 15 (280 mg, 0.485 mmol) and methyl 6-(bromomethyl)nicotinate (0.19 g, 0.825 mmol) gave the title compound (267 mg, 76%) as colorless crystals.

Ethyldiisopropylamine (0.48 ml, 2.76 mmol) and a solution of dimethylaluminium chloride in hexane (0.98 M, 1.69 ml, 1.656 mmol) were added dropwise to a solution of methylamine (2M in THF) (1.38 ml, 2.76 mmol) in dichloromethane (4 ml) under ice cooling, and the mixture was stirred at room temperature for 30 minutes. A solution of the compound obtained in Example 20 (200 mg, 0.275 mmol) in dichloromethane (14 ml) was added to the mixture, and the mixture was stirred at room temperature for 2 days. The reaction mixture was combined with an aqueous solution of sodium hydrogen carbonate and extracted with chloroform. The organic layer was washed with brine, dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by NH-silica gel column chromatography (Fuji Silysia Chemical) (eluent:ethyl acetate/methanol=40/1) and recrystallized from dichloromethane/methanol/diethyl ether to give the title compound (68 mg, 34%) as colorless crystals.

The similar reaction as described in Example 2 by using the compound obtained in Reference Example 15 (578 mg, 1 mmol) and ethyl 6-hydroxymethyl-2-pyridinecarboxylate (797 mg, 4.4 mmol) gave the title compound (590 mg, 80%) as colorless crystals.

The similar reaction as described in Example 21 by using the compound obtained in Example 22 (300 mg, 0.413 mmol) and a solution of methylamine in THF (2 M, 2.07 ml, 4.13 mmol) gave the title compound (158 mg, 53%) as colorless crystals.

The compound obtained in Reference Example 15 (289 mg, 0.5 mmol) was dissolved in DMF (30 ml), and ethyldiisopropylamine (0.44 ml, 2.5 mmol) and 1-bromo-2-chloroethane (0.17 ml, 2.5 mmol) were added thereto. The reaction mixture was stirred at 50-60° C. for 1 hour, combined with an aqueous solution of sodium hydrogen carbonate and extracted with ethyl acetate. The combined organic layer was dried over magnesium sulfate and concentrated under reduced pressure to give a halide. A solution of the obtained halide, imidazole (177 mg, 2.6 mmol) and potassium carbonate (72 mg, 0.52 mmol) in DMF (4 ml) was stirred at room temperature for 18 hours and at 50-60° C. for 1 hour, combined with water and extracted with ethyl acetate. The organic layer was washed with brine, dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by NH-silica gel column chromatography (Fuji Silysia Chemical) (eluent:ethyl acetate/methanol=80/1 to 20/1) and recrystallized from dichloromethane/methanol/diethyl ether to give the title compound (40 mg, 23%) as colorless crystals.

The similar reaction as described in Example 24 by using the compound obtained in Reference Example 15 (289 mg, 0.5 mmol) and 2-(2-hydroxyethyl)imidazole (292 mg, 2.6 mmol) gave the title compound (19 mg, 10%) as colorless crystals.

The similar reaction as described in Example 5 by using the compound obtained in Reference Example 3 (320 mg, 0.5 mmol) and 4-aminophenoxy-N-methylacetamide (135 mg, 0.75 mmol) gave the title compound (192 mg, 51%) as colorless crystals.

The similar reaction as described in Example 5 by using the compound obtained in Reference Example 3 (320 mg, 0.5 mmol) and 4-aminophenyl-N-ethylacetamide (134 mg, 0.75 mmol) gave the title compound (145 mg, 38%) as pale yellow crystals.

The similar reaction as described in Example 5 by using the compound obtained in Reference Example 3 (320 mg, 0.5 mmol) and β-alanine methyl ester (115 mg, 0.75 mmol) gave the title compound (255 mg, 75%) as a pale yellow amorphous compound.

The similar reaction as described in Example 21 by using the compound obtained in Example 28 (300 mg, 0.413 mmol) and a solution of dimethylamine in THF (2M) (1.63 ml, 3.26 mmol) gave the title compound (73 mg, 33%) as pale yellow powders.

The similar reaction as described in Example 5 by using the compound obtained in Example 28 (320 mg, 0.5 mmol) and 4-aminohexanol (86 mg, 0.75 mmol) gave the title compound (154 mg, 45%) as colorless crystals.

Production of N-(4-(1-(2,6-difluorobenzyl)-5-((methyl(2-(1H-tetrazol-1-yl)ethyl)amino)methyl)-2,4-dioxo-3-phenyl-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea (2)

Potassium carbonate (2.075 g, 15 mmol) was added to a solution of tetrazole (0.70 g, 10 mmol) and 1-bromo-2-chloroethane (1.25 ml, 15 mmol) in DMF (5 ml), and the mixture was stirred at room temperature for 16 hours. The reaction mixture was combined with saturated brine and extracted twice with ethyl acetate. The combined extract was dried over magnesium sulfate and concentrated under reduced pressure to give a halide (4.23 g including DMF).

N,N-diisopropylethylamine (1.16 ml, 6.67 mmol) and the halide obtained above were added to a solution of the compound obtained in Reference Example 15 (578 mg, 1 mmol) in DMF (10 ml), and the mixture was stirred at 60-70° C. for 16 hours. The reaction mixture was combined with an aqueous solution of sodium hydrogen carbonate and extracted with ethyl acetate. The organic layer was washed with brine, dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent:ethyl acetate/hexane=9/1) and recrystallized from dichloromethane/methanol/diethyl ether to give the title compound 2-yl form (80 mg, 12%) and l-yl form (38 mg, 57%) as colorless crystals.

The similar reaction as described in Example 5 by using the compound obtained in Reference Example 3 (410 mg, 0.64 mmol) and 2-aminoethanol (0.06 ml, 0.96 mmol) gave the title compound (76 mg, 19%) as colorless crystals.

The similar reaction as described in Example 2 by using the compound obtained in Reference Example 15 (404 mg, 0.7 mmol) and 1-(3-hydroxypropyl)-2-pyrrolidone (0.55 g, 3.85 mmol) gave the title compound (322 mg, 66%) as colorless crystals.

The similar reaction as described in Example 2 by using the compound obtained in Reference Example 15 (404 mg, 0.7 mmol) and 1-(3-hydroxyethyl)-2-pyrrolidone (0.50 g, 3.85 mmol) gave the title compound (290 mg, 60%) as colorless crystals.

The similar reaction as described in Example 2 by using the compound obtained in Reference Example 17 (350 mg, 0.588 mmol) and 1-(3-hydroxyethyl)-2-pyrrolidone (0.50 g, 3.85 mmol) gave the title compound (283 mg, 68%) as pale yellow crystals.

The similar reaction as described in Example 2 by using the compound obtained in Reference Example 17 (350 mg, 0.588 mmol) and 2-methylaminoethanol (0.29 g, 3.85 mmol) gave the title compound (263 mg, 61%) as pale yellow crystals.

The similar reaction as described in Example 2 by using the compound obtained in Reference Example 15 (404 mg, 0.7 mmol) and (L)-2-hydroxymethylpyrrolidine (0.39 g, 3.85 mmol) gave the title compound (262 mg, 51%) as colorless crystals.

The similar reaction as described in Example 2 by using the compound obtained in Reference Example 17 (400 mg, 0.672 mmol) and (L)-2-hydroxymethylpyrrolidine (0.39 g, 3.85 mmol) gave the title compound (222 mg, 44%) as colorless crystals.

The similar reaction as described in Example 2 by using the compound obtained in Reference Example 15 (188 mg, 0.325 mmol) and (R)-2-hydroxymethylpyrrolidine (0.14 g, 1.38 mmol) gave the title compound (136 mg, 57%) as colorless crystals.

Diethyl cyanophosphate (245 mg) and N-ethyldiisopropylamine (284 μl) were added to a solution of the compound obtained in Reference Example 7 (607 mg) and 2-aminopyridine (142 mg) in DMF (10 ml) under ice cooling, and the reaction mixture was warmed gradually to room temperature and stirred for 13 hours. The reaction mixture was distributed between ethyl acetate and water. The organic layer was successively washed with water and saturated brine, dried over magnesium sulfate, and concentrated under reduced pressure. The obtained residue was roughly purified by aminopropyl silica gel column chromatography (Fuji Silysia Chemical). The obtained crude amide (350 mg) was dissolved in ethanol (25.5 ml), and a solution of 28% sodium methoxide in methanol (196 mg) was added thereto. The mixture was stirred at room temperature for 15 hours. The reaction mixture was neutralized with 1N hydrochloric acid (1 ml), and the solvent was distilled off. The residue was distributed between ethyl acetate and water. The organic layer was successively washed with water and saturated brine, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure. The obtained residue was purified by aminopropyl silica gel column chromatography (45 g, eluent:ethyl acetate/hexane 7/3 to ethyl acetate) and recrystallized from THF-ethanol to give the title compound (210 mg) as colorless crystals.

To a solution of the compound obtained in Reference Example 14 (251 mg) in DMF (4.3 ml) were added 2-ethoxyethyl chloride (141 mg), N-ethyldiisopropylamine (245 μl) and potassium iodide (107 mg), and the mixture was stirred at 60° C. for 24 hours. The reaction mixture was distributed between ethyl acetate and water. The organic layer was washed with saturated brine and dried over magnesium sulfate. The solvent was distilled off, and the residue was purified by aminopropyl silica gel column chromatography (Fuji Silysia Chemical) (45 g, eluent: ethyl acetate/hexane=3/2 to 4/1) and recrystallized from ethyl acetate to give the title compound (62 mg) as colorless crystals.

By a similar manner to Example 40, a crude amide (270 mg) was obtained from the compound obtained in Reference Example 7 (304 mg), diethyl cyanophosphate (153 μl), 2-amino-5-fluoropyridine (113 mg) and N-ethyldiisopropylamine (190 μl), and the title compound (113 mg) was obtained from the crude amide by using methanol (19 ml) and a solution of 28% sodium methoxide in methanol (146 mg).

The similar reaction as described in Example 40 by using the compound obtained in Reference Example 7 (304 mg), diethyl cyanophosphate (153 μl), 2-amino-5-bromopyridine (173 mg) and N-ethyldiisopropylamine (190 μl) gave a crude amide (228 mg). Furthermore, the similar reaction by using methanol (14.5 ml) and a solution of 28% sodium methoxide in methanol (112 mg) gave the title compound (113 mg).

The similar reaction as described in Example 40 by using the compound obtained in Reference Example 7 (304 mg), diethyl cyanophosphate (153 μl), 2-amino-5-methylpyridine (109 mg) and N-ethyldiisopropylamine (190 μl) gave a crude amide (188 mg). Furthermore, the similar reaction by using methanol (13.5 ml), a solution of 28% sodium methoxide in methanol (103 mg) gave the title compound (122 mg).

The similar reaction as described in Example 40 by using the compound obtained in Reference Example 7 (423 mg), diethyl cyanophosphate (212 μl), 2-amino-6-methylpyridine (151 mg) and N-ethyldiisopropylamine (265 μl) gave a crude amide (242 mg). Furthermore, the similar reaction by using methanol (17 ml), a solution of 28% sodium methoxide in methanol (131 mg) gave the title compound (145 mg).

The similar reaction as described in Example 40 by using the compound obtained in Reference Example 7 (455 mg), diethyl cyanophosphate (227 μl), 2-amino-3-methoxy-6-methylpyridine (208 mg) and N-ethyldiisopropylamine (310 μl) gave a crude amide (130 mg). Furthermore, the similar reaction by using methanol (8.5 ml), a solution of 28% sodium methoxide in methanol (666 mg) gave the title compound (107 mg).

The similar reaction as described in Example 40 by using the compound obtained in Reference Example 7 (455 mg), diethyl cyanophosphate (285 μl), 2-amino-3-methoxypyridine (233 mg) and N-ethyldiisopropylamine (388 μl) gave a crude amide (226 mg). Furthermore, the similar reaction by using methanol (15.5 ml), a solution of 28% sodium methoxide in methanol (120 mg) gave the title compound (115 mg).

The similar reaction as described in Example 40 by using the compound obtained in Reference Example 7 (1.22 g), diethyl cyanophosphate (456 μl), 4-(1-hydroxy-1-methylethyl)aniline (454 mg) and N-ethyldiisopropylamine (569 μl) gave a crude amide (1.12 g). Furthermore, the similar reaction by using ethanol (60 ml), a solution of 28% sodium methoxide in methanol (579 mg) gave the title compound (849 mg).

The similar reaction as described in Example 40 by using 4-(N-benzyl-N-methylaminomethyl)-2-[N-(2,6-difluorobenzyl)-N-ethoxycarbonylamino]-5-[4-(3-methoxyureido)phenyl]thiophene-3-carboxylic acid (470 mg), diethyl cyanophosphate (167 μl), 4-(1-hydroxy-1-methylethyl)aniline (166 mg) and N-ethyldiisopropylamine (209 μl) gave a crude amide (422 mg). Furthermore, the similar reaction by using ethanol (25.5 ml) and sodium ethoxide (70 mg) gave the title compound (113 mg).

The similar reaction as described in Example 40 by using the compound obtained in Reference Example 7 (364 mg), diethyl cyanophosphate (136 μl), 4-(1-methoxy-1-methylethyl)aniline (149 mg) and N-ethyldiisopropylamine (171 μl) gave a crude amide (292 mg). Furthermore, the similar reaction by using ethanol (19 ml) and a solution of 28% sodium methoxide in methanol (146 mg) gave the title compound (146 mg).

The similar reaction as described in Example 40 by using the compound obtained in Reference Example 7 (455 mg), diethyl cyanophosphate (228 μl), 2-amino-1-methyl-1H-imidazole hydrochloride (214 mg) and N-ethyldiisopropylamine (586 μl) gave a crude amide (48 mg). Furthermore, the similar reaction by using methanol (3.3 ml) and a solution of 28% sodium methoxide in methanol (25 mg) gave the title compound (17 mg).

The similar reaction as described in Example 40 by using the compound obtained in Reference Example 7 (455 mg), diethyl cyanophosphate (228 μl), isoamylamine (139 mg) and N-ethyldiisopropylamine (310 μl) gave a crude amide (102 mg). Furthermore, the stirring with methanol (7 ml), a solution of 28% sodium methoxide in methanol (55 mg) at 55° C. for 20 hours and the similar treatment gave the title compound (80 mg).

The similar reaction as described in Example 40 by using the compound obtained in Reference Example 7 (455 mg), diethyl cyanophosphate (228 μl), 2-methoxyethylamine (120 mg) and N-ethyldiisopropylamine (310 μl) gave a crude amide (266 mg). Furthermore, the similar reaction by using methanol (19.6 ml) and a solution of 28% sodium methoxide in methanol (152 mg) gave the title compound (140 mg).

The similar reaction as described in Example 40 by using the compound obtained in Reference Example 7 (455 mg), diethyl cyanophosphate (228 μl), 2-ethoxyethylamine (143 mg) and N-ethyldiisopropylamine (310 μl) gave a crude amide (259 mg). Furthermore, the similar reaction by using methanol (18.7 ml) and a solution of 28% sodium methoxide in methanol (144 mg) gave the title compound (193 mg).

The similar reaction as described in Example 40 by using the compound obtained in Reference Example 7 (455 mg), diethyl cyanophosphate (228 μl), isopropylamine (95 mg) and N-ethyldiisopropylamine (310 μl) gave a crude amide (306 mg). Furthermore, the stirring with methanol (23.2 ml) and a solution of 28% sodium methoxide in methanol (179 mg) at 60° C. for 17 hours and the similar treatment gave the title compound (192 mg).

The similar reaction as described in Example 40 by using the compound obtained in Reference Example 7 (455 mg), diethyl cyanophosphate (285 μl), 3-amino-6-chloropyridine (243 mg) and N-ethyldiisopropylamine (388 μl) gave a crude amide (207 mg). Furthermore, the similar reaction by using methanol (14.2 ml) and a solution of 28% sodium methoxide in methanol (109 mg) gave the title compound (132 mg).

The similar reaction as described in Example 40 by using the compound obtained in Reference Example 7 (364 mg), diethyl cyanophosphate (182 μl), 3-aminopyridazine hydrochloride (158 mg) and N-ethyldiisopropylamine (414 μl) gave a crude amide (55 mg). Furthermore, the similar reaction by using methanol (4 ml) and a solution of 28% sodium methoxide in methanol (30 mg) gave the title compound (15 mg).

The similar reaction as described in Example 40 by using the compound obtained in Reference Example 7 (607 mg), diethyl cyanophosphate (304 μl), o-methylhydroxylamine hydrochloride (334 mg) and N-ethyldiisopropylamine (1.04 ml) gave a crude product, which was purified by aminopropyl silica gel column chromatography (Fuji Silysia Chemical) to give the title compound (283 mg).

The similar reaction as described in Example 40 by using the compound obtained in Reference Example 7 (607 mg), diethyl cyanophosphate (304 μl), methylamine hydrochloride (270 mg) and N-ethyldiisopropylamine (1.04 ml) gave a crude amide (133 mg). Furthermore, the stirring with methanol (9.1 ml) and a solution of 28% sodium methoxide in methanol (70 mg) at 60° C. for 17 hours and the similar treatment gave the title compound (83 mg).

Into a solution of N-(4-(5-((benzyl(methyl)amino)methyl)-1-(2,6-difluorobenzyl)-1,2,3,4-tetrahydro-2,4-dioxo-3-phenylthieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea (7.7 g, 11.53 mmol) in THF (200 ml) which was cooled with a dry ice-acetone bath was added α-chloroethyl chloroformate (1.7 ml, 11.64 mmol). The temperature of the mixture was elevated up to room temperature, and the mixture was stirred for 2.5 hours. The reaction mixture was diluted with saturated aqueous solution of sodium bicarbonate and extracted with chloroform. The organic layer was washed with saturated aqueous solution of sodium chloride, dried over magnesium sulfate and concentrated under reduced pressure.

The residue was purified with silica gel chromatography (eluent; ethyl acetate/chloroform; from 1/4 to 1/3) and recrystallized from chloroform/diethylether, whereby the title compound (5.66 g, 84%) was obtained as white crystals.

The similar reaction as described in Example 5 by using 4-(N-benzyl-N-methylaminomethyl)-2-(N-(2,6-difluorobenzyl)-N-ethoxycarbonylamino)-5-(4-(3-methoxyureido)phenyl)thiophene-3-carboxylic acid (5.0 g, 7.83 mmol) and p-anisidine (1.93 g, 15.65 mmol) gave N-(4-(5-((benzyl(methyl)amino)methyl)-1-(2,6-difluorobenzyl)-3-(4-methoxyphenyl)-2,4-dioxo-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea (3.39 g, 62%) as white solids. The similar reaction as described in Reference Example 18 by using the compound (80 mg, 0.11 mmol) gave the title compound (50 mg, 74%) as white crystals.

The similar reaction as described in Example 5 by using 4-(N-benzyl-N-methylaminomethyl)-2-(N-(2,6-difluorobenzyl)-N-ethoxycarbonylamino)-5-(4-(3-methoxyureido)phenyl)thiophene-3-carboxylic acid (10.0 g, 15.65 mmol) and 4-aminocyclohexanol (3.61 g, 31.30 mmol) gave N-(4-(5-((benzyl(methyl)amino)methyl)-1-(2,6-difluorobenzyl)-3-(4-hydroxycyclohexyl)-2,4-dioxo-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea (5.20 g, 48%) as white crystals. The similar reaction as described in Reference Example 18 by using the compound (500 mg, 0.72 mmol) gave the title compound (180 mg, 41%) as white crystals.

The similar reaction as described in Example 5 by using 4-(N-benzyl-N-methylaminoethyl)-2-(N-(2,6-difluorobenzyl)-N-ethoxycarbonylamino)-5-(4-(3-methoxyureido)phenyl)thiophene-3-carboxylic acid (10.0 g, 15.65 mmol) and 2-amino-5-fluoropyridine (3.51 g, 31.30 mmol) gave N-(4-(5-((benzyl(methyl)amino)methyl)-1-(2,6-difluorobenzyl)-3-(5-fluoropyridin-2-yl)-2,4-dioxo-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea (3.39 g, 32%) as white solids. The similar reaction as described in Reference Example 18 by using the compound (1.0 g, 1.46 mmol) gave the title compound (560 mg, 64%) as white solids.

The similar reaction as described in Reference Example 14 by using N-(4-(5-((benzyl(methyl)amino)methyl)-1-(2,6-difluorobenzyl)-3-(4-hydroxycyclohexyl)-2,4-dioxo-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea (5.20 g, 48%) obtained in Reference Example gave the title compound (5.20 g, 48%) as white crystals.

A mixture of a solution of methoxyacetone (2.92 g, 33.14 mmol) and methylamine in 2.0 M THF (100 ml, 200.0 mmol) and a solution of acetic acid (0.5 ml) in THF (100 ml) was stirred under ice cooling for 30 minutes. To the mixture was added triacetoxy sodium borohydride (14.05 g, 66.28 mmol), and the mixture was stirred at room temperature for 4 days. To the reaction mixture were added saturated aqueous solution of sodium bicarbonate (100 ml), ethyl acetate ethyl acetate (100 ml) and benzyloxycarbonylchloride (8.48 g, 49.71 mmol), successively, and the mixture was stirred at room temperature for 5 hours. The reaction mixture was distributed between saturated aqueous solution of sodium bicarbonate and ethyl acetate. The ethyl acetate layer was washed with saturated brine and dried over magnesium sulfate, and the solvent was distilled off. The residue was purified by silica gel chromatography (eluent:ethyl acetate/hexane; from 6/1 to 4/1) to obtain colorless liquid (4.24 g, 54%). A mixed solution of the colorless liquid (0.94 g, 3.96 mmol) and 10% palladium-carbon (94 mg) in ethanol (10 ml) was stirred at room temperature under hydrogen atmosphere for 2 hours, and subjected to filtration. The filtrate was concentrated under reduced pressure to give the title compound (220 mg, 54%) as pale yellow liquid.

The similar reaction as described in Reference Example 26 by using compound (100 mg, 0.17 mmol) obtained in Reference Example 18 and (S)-(−)—N-α-dimethylbenzylamine (28 mg, 0.21 mmol) gave the title compound (61 mg, 53%) as white crystals.

The similar reaction as described in Reference Example 26 by using compound (100 mg, 0.17 mmol) obtained in Reference Example 18 and (R)-(+)-1-phenylethylamine (25 mg, 0.21 mmol) gave the title compound (56 mg, 49%) as white crystals.

The similar reaction as described in Reference Example 26 by using the compound (100 mg, 0.17 mmol) obtained in Reference Example 18 and (S)-(−)-1-phenylethylamine (25 mg, 0.21 mmol) gave the title compound (56 mg, 49%) as white crystals.

Under ice cooling, to a solution of 1-phenyl-1-propanol (1.0 g, 7.34 mmol) and N-ethyldiisopropylamine (1.42 g, 11.01 mmol) in dichioromethane (10 ml) was added methanesulfonyl chloride (690 μl, 8.81 mmol), and the mixture was stirred at room temperature for 5 hours. The reaction mixture was diluted with saturated aqueous solution of sodium bicarbonate and extracted with ethyl acetate. The extract was washed with saturated brine, dried over magnesium sulfate, and solvent was distilled off under reduced pressure. The residue was purified by silica gel chromatography (eluent:ethyl acetate/hexane:5/1) to give 1-chloro-1-phenylpropane (610 mg, 54%) as pale yellow liquid.

The similar reaction as described in Reference Example 26 by using the compound (100 mg, 0.17 mmol) obtained in Reference Example 15 and the 1-chloro-1-phenylpropane (32 mg, 0.21 mmol) obtained above gave the title compound (45 mg, 38%) as white crystals.

The similar reaction as described in Example 20 by using the compound (150 mg, 0.17 mmol) obtained in Reference Example 15 and α-bromophenylmethyl acetate (71 mg, 0.31 mmol) gave the title compound (73 mg, 39%) as white crystals.

A solution of methylphenethylether (1.0 g, 7.34 mmol), NBS (1.96 g, 11.01 mmol) and AIBN (240 mg, 1.47 mmol) in carbon tetrachloride (30 ml) was heated under reflux for 2 hours. After cooling, the reaction mixture was diluted with saturated aqueous solution of sodium bicarbonate and extracted with chloroform. The extract was washed with saturated brine, dried over magnesium sulfate and concentrated under reduced pressure to give a crude 2-methoxy-1-phenylbromoethane (1.33 g, 84%) as brown liquid.

The similar reaction as described in Example 14 by using the compound (200 mg, 0.34 mmol) obtained in Reference Example 15 and the crude bromide (50 mg) obtained above gave the title compound (42 mg, 18%) as white crystals.

The similar reaction as described in Example 14 by using the compound (160 mg, 0.26 mmol) obtained in Reference Example 15 and the above bromide (62 mg, 0.33 mmol) obtained above gave the title compound (133 mg, 75%) as white crystals.

The title compound (40 mg) was optically resolved by a preparative HPLC by using CHIRALPAK AD (50 mmI.D.×500 mmL), in which mobile phase is hexane/2-propanol(=3/2), to obtain 19 mg of an optical isomer having an retention time of 25 minutes (99.9% ee) and 19 mg of another isomer having retention time of 29 minutes (99.0% ee) in an analysis using CHIRALPAK AD (4.6 mmI.D.×250 mmL), both of which are white powders.

The similar reaction as described in Reference Example 26 by using the compound (100 mg, 0.17 mmol) obtained in Reference Example 18 and the compound (21 mg, 0.21 mmol) obtained in Reference Example 24 gave the title compound (24 mg, 21%) as white crystals.

The title compound (48 mg) was optically resolved by preparative HPLC by using CHIRALPAK AD(50 mmI.D.×500 mmL), in which mobile phase is hexane/2-propanol (65/35), to obtain 22 mg of an optical isomer having an retention time of 49 minutes (99.0% ee) and 21 mg of another isomer having retention time of 54 minutes (99.0% ee) in an analysis using CHIRALPAK OD (4.6 mmI.D.×250 mmL), both of which are white powders.

The title compound (20 mg) was optically resolved by a preparative HPLC by using CHIRALPAK AD (50 mmI.D.×500 mmL), in which mobile phase is hexane/2-propanol (1/1), to obtain 10 mg of an optical isomer having an retention time of 23 minutes (99.9% ee) and 11 mg of another isomer having retention time of 28 minutes (99.2% ee) in an analysis using CHIRALPAK AD (4.6 mmI.D.×250 mmL), each of which is colorless oily substance.

The similar reaction as described in Example 4 by using the compound (800 mg, 1.24 mmol) obtained in Reference Example 25 and 2-bromoethylethylether (207 mg, 1.49 mmol) gave the title compound (407 mg, 58%) as white solids.

The mixture was extracted with ethyl acetate. The organic layer was washed with brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure to give the title compound (2.98 g, quant.) as colorless oily substance.

To a solution of compound (2.96 g, 10 mmol) obtained in Reference Example 42 in DMF (10 ml) was added potassium phthalimide (1.85 g, 10 mmol). The reaction liquid was stirred at 140° C. for 30 hours, and water was added. The mixture was extracted with ethyl acetate. The organic layer was washed with brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent;ethyl acetate/hexane=1/10) to give the title compound (2.79 g, 80%) as pale yellow oily substance.

A mixture of compound (1.395 g, 4.0 mmol) obtained in Reference Example 43, concentrated hydrochloric acid (10 ml), acetic acid (7 ml) and water (10 ml) was refluxed for 24 hours. The reaction liquid was concentrated and water was added. The precipitated phthalic acid was filtered off and filtrate was concentrated. The residue was recrystallized from ethyl acetate to give the title compound (257.2 mg, 46%) as pale purple crystals.

To a solution of lithium aluminum hydride (9.34 g, 246 mmol) in THF (150 ml) was dropwise added a solution of dimethylcyclopropane-1,1-dicarboxylate (25.95 g, 164.1 mmol) in THF (150 ml) under ice cooling. The solution was stirred at 0° C. for 2 hours, and to the solution were slowly added water (9.5 ml), 15% aqueous solution of NaOH (9.5 ml) and water (30 ml), successively. Insoluble matters were filtered off, and the filtrate was concentrated to give cyclopropane-1,1-dimethylmethanol as colorless oily substance.

To a solution of the above oily substance in 1,2-dimethoxyethane (150 ml) was added sodium hydride (60% oil, 6.56 g, 164 mmol), and the mixture was stirred at room temperature for 1 hour. To the reaction liquid was dropwise added a solution of tert-butylchlorodimethylsilane (24.7 g, 164 mmol) in 1,2-dimethoxyethane (100 ml), and the mixture was stirred at room temperature over night. The reaction liquid was poured into water and extracted twice with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by silicagel column chromatography (eluent;ethyl acetate/hexane=1/15-1/9) to give the title compound (28.19 g, 79%) as colorless oily substance.

To a solution of compound (2.0 g, 5.79 mmol) obtained in Reference Example 47 in ethanol (20 ml) was added hydrazine monohydrate (0.42 ml, 8.68 mmol) and the mixture was refluxed for 2 hours. Insoluble matters were filtered off and the filtrate was concentrated. The residue was distributed between ethyl acetate and aqueous solution of 1N-sodium hydroxide, and extracted with ethyl acetate. The organic layer was washed with brine, dried over magnesium sulfate and concentrated to give the title compound (1.15 g, 92%) as pale yellow oily substance.

The similar reaction as described in Reference Example 18 by using the N-(4-(1-(2,6-difluorobenzyl)-5-(((2-methoxyethyl)(methyl)amino)methyl)-3-(6-methoxy-3-pyridinyl)-2,4-dioxo-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea (49.3 mg, 0.074 mmol) gave the title compound (44.1 mg, quant.) as white powders.

The similar reaction as described in Reference Example 18 by using the N-(4-(1-(2,6-difluorobenzyl)-5-(((2-methoxyethyl)(methyl)amino)methyl)-3-(6-methoxy-3-pyridazinyl)-2,4-dioxo-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea (1.34 g, 2 mmol) gave the title compound (888.5 mg, 71%) as pale yellow powders.

The similar reaction as described in Reference Example 14 by using the compound (1.75 g, 2.54 mmol) obtained in Example 69 described later gave the title compound (1.60 g, quant.) as colorless crystals.

The similar reaction as described in Reference Example 26 by using the compound (130 mg, 0.192 mmol) obtained in Reference Example 19 described later and a solution of dimethylamine in THF (0.96 ml, 1.92 mmol) gave the title compound (43.4 mg, 36%) as colorless crystals.

The similar reaction as described in Reference Example 26 by using the compound (110 mg, 0.175 mmol) obtained in Reference Example 50 and pyrrolidine (124 mg, 1.75 mmol) gave the title compound (68.1 mg, 60%) as colorless crystals.

The similar reaction as described in Reference Example 26 by using the compound (110 mg, 0.175 mmol) obtained in Reference Example 50 and morpholine (152 mg, 1.75 mmol) gave the title compound (78.0 mg, 67%) as colorless crystals.

The similar reaction as described in Reference Example 26 by using the compound (120 mg, 0.191 mmol) obtained in Reference Example 50 and N-benzyl-N-(2-methoxyethyl)amine (316 mg, 1.91 mmol) gave the title compound (105.1 mg, 74%) as white powders.

The similar reaction as described in Example 5 by using the compound (1.82 g, 3 mmol) obtained in Reference Example 7 and 1-amino-2-propanol (0.45 g, 6 mmol) gave the title compound (1.11 g, 60%) as white powders.

To a solution of oxalyl chloride (0.20 g, 1.58 mmol) in dichloromethane (2 ml) was dropwise added a solution of dimethylsulfoxide (163 mg, 2.09 mmol) in dichloromethane (2 ml) under cooling at −78° C. After stirring for 10 minutes, a solution of the compound (308 mg, 0.5 mmol) obtained in Example 61 in dichloromethane (2 ml) was dropwise added. After stirring for further 30 minutes, triethylamine (0.40 ml, 2.88 mmol) was dropwise added, and the mixture was stirred at 0° C. for 2 hours. To the mixture was added an aqueous solution of ammonium chloride at 0° C., and the mixture was extracted with ethyl acetate. The organic layer was washed with brine, dried over anhydrous magnesium sulfate, and concentrated with under reduced pressure. The residue was purified by NH-silica gel (Produced by Fuji Silysia Chemical Ltd.) column chromatography (eluent;ethyl acetate), and the obtained powders was washed with diisopropyl ether and hexane to give the title compound (40.7 mg, 13%) as pale yellow powders.

The similar reaction as described in Example 5 by using the compound (303 mg, 0.5 mmol) obtained in Reference Example 7 and 3,3-dimethyl-2-oxobutylamine hydrochloride (U.S. Pat. No. 6,096,688) (152 mg, 1 mmol) gave the title compound (195.7 mg, 60%) as pale yellow crystals.

The similar reaction as described in Example 5 by using the compound (303 mg, 0.5 mmol) obtained in Reference Example 7 and 2,2,2-trifluoroethylamine (99 mg, 1 mmol) gave the title compound (52.9 mg, 16%) as colorless crystals.

To a mixed solution of compound (120 mg, 0.182 mmol) obtained in Example 63 in methanol (6 ml) and THF (4 ml) was added sodium borohydride (6.9 mg, 0.182 mmol) under ice cooling. The reaction liquid was stirred at room temperature for 1.5 hours and concentrated under reduced pressure. The residue was distributed between water and ethyl acetate, and the organic layer was extracted. The extract was washed with brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by NH-silica gel (Produced by Fuji Silysia Chemical Ltd.) column chromatography (eluent;ethyl acetate), and recrystallized from ethyl acetate and diethylether to give the title compound (81.1 mg, 68%) as colorless crystals.

The similar reaction as described in Example 5 by using the compound (303 mg, 0.5 mmol) obtained in Reference Example 7 and compound (89 mg, 1 mmol) obtained in Reference Example 41 gave the title compound (133.9 mg, 42%) as colorless crystals.

The similar reaction as described in Example 5 by using the compound (303 mg, 0.5 mmol) obtained in Reference Example 7 and compound (140 mg, 1 mmol) obtained in Reference Example 44 gave the title compound (36.8 mg, 11%) as pale yellow crystals.

The similar reaction as described in Example 5 by using the compound (606 mg, 1 mmol) obtained in Reference Example 7 and compound (431 mg, 2 mmol) obtained in Reference Example 47 gave the thienopyrimidine cyclized form (549.3 mg, 72%). The above cyclized form was dissolved in THF (10 ml), and a solution of tetrabutylammoniumfluoride in 1M THF (1.7 ml, 1.7 mmol) was added. The mixture was stirred at room temperature for 20 hours. The reaction liquid was distributed between water and ethyl acetate, and the organic layer was extracted. The organic layer was washed with brine, dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by NH-silica gel (Produced by Fuji Silysia Chemical Ltd.) column chromatography (eluent;ethyl acetate), and recrystallized from ethyl acetate and diethylether to give the title compound (196.1 mg, 43%) as colorless crystals.

The similar reaction as described in Example 5 by using 4-(N-benzyl-N-methylaminomethyl)-2-[N-(2,6-difluorobenzyl)-N-ethoxycarbonylamino]-5-[4-(3-methoxyureido)phenyl]thiophene-3-carboxylic acid (3.19 g, 5 mmol) and trans-4-aminocyclohexanol (1.44 g, 12.5 mmol) gave the title compound (1.80 g, 52%) as colorless crystals.

The similar reaction as described in Example 4 by using the compound (300 mg, 0.5 mmol) obtained in Reference Example 51 and 2-bromoethylmethylether (0.69 g, 5 mmol) gave the title compound (75.1 mg, 23%) as colorless crystals.

The similar reaction as described in Example 5 by using the compound (303 mg, 0.5 mmol) obtained in Reference Example 7 and 3-amino-6-methylpyridazine (136 mg, 1.25 mmol) gave the title compound (48.3 mg, 15%) as colorless crystals.

The similar reaction as described in Example 5 by using 4-(N-benzyl-N-methylaminomethyl)-2-[N-(2,6-difluorobenzyl)-N-ethoxycarbonylamino]-5-[4-(3-methoxyureido)phenyl]thiophene-3-carboxylic acid (639 mg, 1 mmol) and 1-amino-2-propanol (0.19 g, 2.5 mmol) gave the title compound (409.7 mg, 63%) as colorless crystals.

The similar reaction as described in Example 5 by using the 4-(N-benzyl-N-methylaminomethyl)-2-[N-(2,6-difluorobenzyl)-N-ethoxycarbonylamino]-5-[4-(3-methoxyureido)phenyl]thiophene-3-carboxylic acid (639 mg, 1 mmol) and compound (0.22 g, 2.5 nimol) obtained in Reference Example 41 gave the title compound (360.9 mg, 54%) as colorless crystals.

The similar reaction as described in Example 2 by using the compound (260 mg, 0.465 mmol) obtained in Reference Example 52 and 2-(2-hydroxyethyl)pyridine (400 mg, 1.63 mmol) gave the title compound (195.9 mg, 63%) as colorless crystals.

The similar reaction as described in Example 2 by using the compound (220 mg, 0.384 mmol) obtained in Reference Example 53 and 2-(2-hydroxyethyl)pyridine (800 mg, 3.25 mmol) gave the title compound (138.2 mg, 53%) as colorless crystals.

The similar reaction as described in Example 5 by using the compound (303 mg, 0.5 mmol) obtained in Reference Example 7 and aminopyrazine (119 mg, 1.25 mmol) gave the title compound (35.2 mg, 11%) as colorless crystals.

The similar reaction as described in Example 5 by using 4-(N-benzyl-N-methylaminomethyl)-2-[N-(2,6-difluorobenzyl)-N-ethoxycarbonylamino]-5-[4-(3-methoxyureido)phenyl]thiophene-3-carboxylic acid (1.28 g, 2 mmol) and 6-chloro-3-aminopyridazine (648 mg, 5 mmol) gave the title compound (0.36 g, 26%) as colorless crystals.

The similar reaction as described in Example 2 by using the compound (135 mg, 0.221 mmol) obtained in Reference Example 54 and 2-(2-hydroxyethyl)pyridine (272 mg, 2.21 mmol) gave the title compound (79.6 mg, 50%) as colorless crystals.

The similar reaction as described in Example 2 by using the compound (800 mg, 1.24 mmol) obtained in Reference Example 25 and 2-(2-hydroxyethyl)pyridine (300 mg, 1.49 mmol) gave the title compound (407 mg, 58%) as white solids.

To a solution of the compound (1.34 g, 2 mmol) obtained in Example 57 in THF (30 ml) was added a solution of 4N HCl in acetic acid (2 ml, 8 mmol), and the mixture was stirred at room temperature for 20 hours, at 50° C. for 10 hours and further at 60° C. for 3 hours. The reaction liquid was distributed between aqueous solution of sodium bicarbonate and ethyl acetate, and the organic layer was extracted. The aqueous layer was subjected to salting-out, and extracted with ethyl acetate. The organic layers were collected and combined and the organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by NH-silica gel (Produced by Fuji Silysia Chemical Ltd.) column chromatography (eluent;ethyl acetate/methanol=8/1). To the eluate was added diisopropylether to give powders. The powders were collected by filtration, washed with diisopropylether to obtain the title compound (539 mg, 41%) as a pale yellow powder.

The similar reaction as described in Example 4 by using the compound (305 mg, 0.5 mmol) obtained in Reference Example 54 and 2-bromoethanol (0.62 g, 5 mmol) gave the title compound (145.7 mg, 45%) as colorless crystals.

The similar reaction as described in Example 4 by using the compound (123 mg, 0.191 mmol) obtained in Reference Example 25 and 2-bromoethanol (239 mg, 1.91 mmol) gave the title compound (47.1 mg, 39%) as colorless crystals.

The similar reaction as described in Example 4 by using the compound (100 mg, 0.164 mmol) obtained in Reference Example 54 and methyl iodide (0.010 ml, 0.164 mmol) gave the title compound (17.3 mg, 17%) as colorless crystals.

The similar reaction as described in Reference Example 26 by using the compound (41.1 mg, 0.067 mmol) obtained in Reference Example 49 and a solution of dimethylamine in THF (0.67 ml, 1.34 mmol) gave the title compound (18.4 mg, 44%) as white powders.

The compound obtained in Example 40 (5 g) was dissolved in distilled water for injection to make the total volume to 100 ml. The solution was subjected to sterile filtration by using membrane filter having a diameter of 0.22 μm (Produced by Sumitomo Electric Industries, Ltd. or Sartorius K.K.), 2 ml each of the solution is poured into a sterilized vial, and is freeze dried in a conventional manner to give freeze-dried injectable preparation.

The compound obtained in Example 41 (5 g) was dissolved in distilled water for injection to make the total volume to 100 ml. The solution was subjected to sterile filtration by using membrane filter having a diameter of 0.22 μm (Produced by Sumitomo Electric Industries, Ltd. or Sartorius K.K.), 2 ml each of the solution is poured into a sterilized vial, and is freeze dried in a conventional manner to give freeze-dried injectable preparation.

Preparation 5

(1)

Compound obtained in Example 40 or Example 41

5

g

(2)

lactose · crystalline cellulose (grain)

330

g

(3)

D-mannitol

29

g

(4)

low substituted hydroxypropylcellulose

20

g

(5)

talc

25

g

(6)

hydroxypropylcellulose

50

g

(7)

aspartame

3

g

(8)

dipotassium glycyrrhizinate

3

g

(9)

hydroxypropylmethylcellulose 2910

30

g

(10)

titanium oxide

3.5

g

(11)

ferric oxide yellow

0.5

g

(12)

light silicic acid anhydride

1

g

The above (1), (3), (4), (5), (6), (7) and (8) are suspended or dissolved in purified water, nuclear particles of (2) are coated with the solution to prepare crude fine grains. The crude fine grains are coated with (9) to (11) to prepare coated fine grains. They are mixed with (12) to prepare 1% KM05283 fine grains (500 g). 500 mg each of the fine grains are wrapped, separately.

Experimental Example 1

(1) Preparation of 125I-Leuprorelin

To a tube containing 10 μl of a 3×10−4 M aqueous solution of leuprorelin and 10 μl of 0.01 mg/ml lactoperoxidase was added 10 μl (37 MBq) of a solution of Na125I. After stirring, 10 μl of 0.001% H2O2 was added, and a reaction was carried out at room temperature for 20 minutes. By adding 700 μl of a 0.05% TFA solution, the reaction was stopped, followed by purification by reversed-phase HPLC. The HPLC conditions used are shown below. 125I-leuprorelin was eluted at a retention time of 26 to 27 minutes.

Monkey GnRH receptor-expressing CHO cells (109 cells) were suspended in phosphate-buffered saline supplemented with 5 mM EDTA (PBS-EDTA) and centrifuged at 100×g for 5 minutes. To the cell pellet, 10 ml of a cell homogenate buffer (10 mM NaHCO3, 5 mM EDTA, pH 7.5) was added, followed by homogenization using the Polytron homogenizer. After centrifugation at 400×g for 15 minutes, the supernatant was transferred to an ultracentrifugation tube and centrifuged at 100,000×g for 1 hour to yield a membrane fraction precipitate. This precipitate was suspended in 2 ml of an assay buffer and centrifuged at 100,000×g for 1 hour. The membrane fraction recovered as a precipitate was again suspended in 20 ml of the assay buffer, dispensed, and stored at −80° C. before use upon thawing.

Human GnRH receptor-expressing CHO cells (109 cells) were suspended in phosphate-buffered saline supplemented with 5 mM EDTA (PBS-EDTA) and centrifuged at 100×g for 5 minutes. To the cell pellet, 10 ml of a cell homogenate buffer (10 mM NaHCO3, 5 mM EDTA, pH 7.5) was added, followed by homogenization using the Polytron homogenizer. After centrifugation at 400×g for 15 minutes, the supernatant was transferred to an ultracentrifugation tube and centrifuged at 100,000×g for 1 hour to yield a membrane fraction precipitate. This precipitate was suspended in 2 ml of an assay buffer and centrifuged at 100,000×g for 1 hour. The membrane fraction recovered as a precipitate was again suspended in 20 ml of the assay buffer, dispensed, and stored at −80° C. before use upon thawing.

(4) Determination of 125I-leuprorelin Binding Inhibition rate

The monkey and human membrane fractions prepared in the above (2) and (3) were diluted with the assay buffer to yield a 200 μg/ml dilution, each of which was then dispensed at 188 μl per tube. To a tube containing the cell membrane fraction of the CHO with monkey GnRH receptors expressed were added 2 μl of a solution of 20 mM compound in 60% DMSO and 10 μl of 38 nM 125I-leuprorelin. To a tube containing the cell membrane fraction of the CHO with human GnRH receptors expressed were added 2 μl of a solution of 2 mM compound in 60% DMSO and 10 μl of 38 nM 125I-leuprorelin. To determine maximum binding quantity, a reaction mixture containing 2 μl of 60% DMSO and 10 μl of 38 nM 125I-leuprorelin was prepared. To determine non-specific binding amount, a reaction mixture containing 2 μl of a solution of 100 μM leuprorelin in 60% DMSO and 10 μl of 38 nM 125I-leuprorelin was prepared.

Where the membrane fraction of the CHO with monkey and human GnRH receptors expressed was used, the reaction was carried out at 25° C. for 60 minutes. After each reaction, the reaction mixture was aspirated and filtered through a polyethyleneimine-treated Whatman glass filter (GF-F). After this filtration, the radioactivity of 125I-leuprorelin remaining on the filter paper was measured with a γ-counter.

The expression (TB-SB)/(TB-NSB)×100 (where SB=radioactivity with the compound added, TB=maximum bound radioactivity, NSB=nonspecifically bound radioactivity) was calculated to find the binding inhibition rate of each test compound. Furthermore, the inhibition rate was determined by varying the concentration of the test substance and the 50% inhibitory concentration (IC50 value) of the compound was calculated from Hill plot. The results are shown in below.

TABLE 1

IC50 value (μM)

Test Compound

Monkey

Human

Ex. Compd. No. 40

0.009

0.0002

Ex. Compd. No. 41

0.003

0.0001

INDUSTRIAL APPLICABILITY

The compound of the present invention possesses excellent gonadotropin-releasing hormone antagonizing activity. It is also good in oral absorbability and excellent in stability and pharmacokinetics. With low toxicity, it is also excellent in safety. Therefore, for example, the compound of the present invention can be used as a preventing or treating agent for hormone-dependent diseases. Concretely, for example, it is effective as a preventing or treating agent for sex hormone-dependent cancers (e.g., prostatic cancer, uterine cancer, breast cancer, pituitary tumor, and the like), prostatic hypertrophy, hysteromyoma, endometriosis, metrofibroma, precocious puberty, amenorrhea syndrome, premenstrual syndrome, multilocular ovary syndrome, polycystic ovary syndrome, acne, alopecia, Alzheimer's disease, and the like; as a pregnancy regulator (e.g., contraceptive, etc.), infertility remedy or menstruation regulator, as a preventing or treating agent for irritable bowel syndrome; or as a preventing agent for postoperative recurrence of sex hormone-dependent cancer, as medicament. It is also effective as an animal estrous regulator, food meat quality improving agent or animal growth regulator in the field of animal husbandry, and as a fish spawning promoter in the field of fishery.

Claims (2)

1. N-(4-(1-(2,6-difluorobenzyl)-5-(((2-ethoxyethyl)(methyl)amino)methyl)-2,4-dioxo-3-(2-pyridinyl)-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea or a salt thereof.

2. N-(4-(1-(2,6-difluorobenzyl)-5-((dimethylamino)methyl)-3-(6-methoxy-3-pyridazinyl)-2,4-dioxo-1,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-6-yl)phenyl)-N′-methoxyurea or a salt thereof.